Method of synthesizing camptothecin-relating compounds

ABSTRACT

The present invention is to prepare efficiently 2′-amino-5′-hydroxypropiophenone corresponding to the AB-ring part of camptothecin (CPT) skeleton and a tricyclic ketone corresponding to the CDE-ring part in order to provide efficiently CPT by the total synthesis, which is a starting material for irinotecan hydrochloride and various kinds of camptothecin derivatives, and to provide stably CPT and its derivatives.

RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No. 10/467,987, filed Feb. 21, 2002, now issued as U.S. Pat. No. 7,126,000, which is a national stage application under 35 U.S.C. § 371 of International Application No. PCT/JP02/01538 designating the United States of America, filed Feb. 1, 2002, the entire disclosures of which are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to a process for synthesizing camptothecin related compound(s). More particularly, the invention relates to a process for preparing intermediates related to the synthesis of camptothecin analogs having an anti-tumor activity and use of said intermediates, and relates to a total synthesis of camptothecin analogs.

BACKGROUND ART

Camptothecin (hereinafter described as CPT) isolated from the bark, root, fruit, leaf and the like of Camtotheca acuminata of Chinese origin is a pentacyclic alkaloid and is known to show the anti-tumor activity by inhibition of a nucleic acid synthesis. In the meantime, as to a camptothecin derivative the induction of diarrhea and the like as a side effect are reported (Gann to Kagaku Ryohou 17, p115-120, 1990), leaving a problem to cause disorder for the gastrointestinal tract, and therefore, various kinds of derivatives have been examined to reduce the toxicity, to increase the effect, and so on.

Thus, the inventors already reported 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin.hydrochloride.trihydrate (hereinafter described as CPT-11), the water soluble semisynthetic derivative of CPT, as a compound which is reduce in toxicity compared to CPT, and it is at present widely used as the anti-tumor agent (general name; irinotecan hydrochloride).

Camptothecin analogs such as CPT-11 can be derived by a chemical modification of CPT obtained from natural materials.

However, owing to an extremely low amount of CPT obtained from natural materials such as Camtotheca acuminata which is the starting material, it is anticipated that according to an increased demand of CPT-11 which is a useful derivative and the like, a sufficient supply of CPT becomes difficult notwithstanding a measure for the starting material supply such as afforestation. Although the total synthesis is also examined, it is the present situation that it has not yet been into practical use.

As a process by total synthesis is known the method of Shen, W. et al. represented by the below reaction scheme via Friedländer reaction of the aminopropiophenone and the tricyclic ketone (J. Org. Chem. 1993, 58, 611-617 “Concise Total Syntheses of dl-Camptothecin and Related Anticancer Drugs.”, though there are problems that the steps are tedious, the yields are not sufficient and only the racemate is synthesized.

In the meantime, although Curran, D. P. et al. carried out a total synthesis by the method using a cascade radical cyclization of the aryl isonitrile and the iodopyridone represented by the below reaction scheme (Chem. Eur. J. 1998, 4, 67-83 “A General Synthetic Approach to the (20S)-Camptothecin Family of Antitumor Agents by a Regiocontrolled Cascade Radical Cyclization of Aryl Isonitriles.”, problems are pointed out in which the yield of the cyclization reaction is not sufficient and deprotection of the protective group is necessary after cyclization.

Additionally, although the above Curran, D. P. et al. synthesized 4-iodo-2-methoxy-6-trimethylsilylpyridine-3-carbaldehyde, an intermediate in the synthesis of the tricyclic ketone part of CPT analogs, according to the below scheme,

this method is highly dangerous due to the necessity to use t-BuLi easily flammable in a large amount industrially, and the reaction at −78° C. as a reaction temperature is required, making it impossible to enlarge the batch size. Further, owing to the necessity of a complicated temperature control in the total reaction system it was not an industrially practical reaction system.

DISCLOSURE OF INVENTION

It is an object of the invention to provide efficiently CPT, which is a starting material for irinotecan hydrochloride and various kinds of camptothecin derivatives, and camptothecin analogs such as 7-ethyl-10-hydroxycamptothecin, which is a key intermediate of the irinotecan hydrochloride synthesis, by a practical total synthesis. Particularly, it is an object of the invention to synthesize an intermediate corresponding to the AB-ring part of camptothecin skeleton and an intermediate corresponding to the CDE-ring part respectively, and further to synthesize camptothecin analogs using these intermediates.

MODE FOR CARRYING OUT THE INVENTION

In view of these circumstances the inventors made an extensive research, and consequently as to the AB-ring part, made Compound (a) (5-hydroxy-2-nitrobenzaldehyde):

a starting material, and found a means to provide CPT and its derivatives stably by an efficient preparation of 2′-amino-5′-hydroxypropiophenone corresponding to the AB-ring part of CPT skeleton, and as to the CDE-ring part starting from Compound (k) (2-methoxy-6-trimethylsilylpyridine (MTP)):

(wherein TMS represents a trimethylsilyl group, and Me represents a methyl group.) found a means to provide CPT and its derivatives stably by an efficient preparation of a tricyclic ketone corresponding to the CDE-ring part of CPT skeleton, and established a total synthetic process for CPT analogs by an appropriate combination of these means without using natural materials, finishing the invention.

Namely, the invention relates to a process for preparing 2′-amino-5′-hydroxypropiophenone corresponding to the AB-ring of CPT skeleton according to the route;

(wherein R represents a protective group.), and relates to a total synthetic process of CPT analogs by the appropriate combination of a process for the tricyclic ketone corresponding to the CDE-ring part of CPT skeleton comprising particularly synthesis of 3-formyl-4-iodo-2-methoxy-6-trimethylsilylpyridine (Compound (l)) from 2-methoxy-6-trimethylsilylpyridine (Compound (k)) or 3-hydroxymethyl-4-iodo-2-methoxy-6-trimethylsilylpyridine (Compound (v)) by improving and optimizing a process according to the synthetic route;

(wherein TMS is a trimethylsilyl group, Me is a methyl group, Et is an ethyl group, Pr is a propyl group, and ^(t)Bu is a t-butyl group.), established on the basis of Curran route (Josien, H,; Ko, S. B.; Bom, D.: Curran, D. P. Chem. Eur. J. 1998, 4 67-83) and Pharmacia & Upjohn route (hereinafter described as P&U route; Heneger, K. E.; Ashford, S, W.; Baughman, T. A.; Sih, J. C.; Gu, R. L. J. Org. Chem. 1997, 62, 6588-6597.) which are synthetic routes currently known. Further, since Compound (v) is a byproduct arising in the process to synthesize 3-(2-butenyloxymethyl)-4-iodo-2-methoxy-6-trimethylsilylpyridine (Compound (m)), in the above synthetic route Compound (l) is described in the downstream.

Particularly, the invention relates to a process for preparing 2′-amino-5′-hydroxypropiophenone to synthesize camptothecin analogs, wherein from Compound (a):

Compound (b):

is produced; and from Compound (b) Compound (c):

is produced; and from Compound (c) Compound (d):

is produced; and from Compound (d) Compound (e):

is produced; wherein R is a protective group which can be deprotected by a catalytic reduction.

Also, the invention relates to the above process, wherein the protective group which can be deprotected by a catalytic reduction is a benzyl group.

Further, the invention relates to the above process, wherein it contains one or more steps selected from the group consisting of

-   (1) a step to obtain Compound (b) mixing Compound (a), a benzylation     reagent and a base, and stirring said mixture in solvent under     reflux; -   (2) a step to obtain Compound (c) by dropping Grignard reagent to     Compound (b) under an inert gas atmosphere; -   (3) a step to obtain Compound (d) mixing Compound (c) and an     oxidizing agent and stirring the mixture; -   (4) a step to obtain Compound (e) by a catalytic reduction of     Compound (d).

Further, the invention relates to the above process wherein in the step (1) the solvent is dimethylformamide.

The invention also relates to the above process wherein in the step (2) the Grignard reagent is vinyl magnesium bromide.

Further, the invention relates to the above process wherein in the step (3) the oxidizing agent is Jones reagent, manganese dioxide or TEMPO-(2,2,6,6-tetramethylpiperidine-1-oxyl)-sodium hypochlorite.

Also, the invention relates to compound represented by formula (c′):

(wherein Bn is a benzyl group.).

Further, the invention relates to compound represented by formula (d′):

(wherein Bn is a benzyl group.).

Also, the invention is a process for preparing 2′-amino-5′-hydroxypropiophenone to synthesize camptothecin analogs, wherein from Compound (a):

Compound (c″):

is produced; and from Compound (c″) Compound (d″):

is produced; and from Compound (d″) Compound (e):

is produced.

Further, the invention relates to the above process, wherein it contains one or more steps selected from the group consisting of

-   (1) a step to obtain Compound (c″) by dropping Grignard reagent to     Compound (a) under an inert gas atmosphere; -   (2) a step to obtain Compound (d″) mixing the Compound (c″) and an     oxidizing agent and stirring the mixture; and -   (3) a step to obtain Compound (e) by a catalytic reduction of     Compound (d″).

The invention also relates to the above process wherein in the step (1) the Grignard reagent is vinyl magnesium bromide.

Further, the invention relates to the above process wherein in the step (2) the oxidizing agent is Jones reagent, manganese dioxide or TEMPO-sodium hypochlorite.

The invention also relates to use of 2′-amino-5′-hydroxypropiophenone, which is obtained by the above process, to the preparation of camptothecin analogs.

Further, the invention relates to a process for preparing camptothecin analogs, comprising reaction of 2′-amino-5′-hydroxypropiophenone obtained by the above process and a tricyclic ketone.

The invention also is a process for preparing the tricyclic ketone to synthesize camptothecin analogs, wherein from Compound (k):

(wherein TMS is a trimethylsilyl group, and Me is a methyl group.), or Compound (v):

(wherein TMS is a trimethylsilyl group, and Me is a methyl group. Compound (l):

(wherein TMS is a trimethylsilyl group, and Me is a methyl group.) is produced; and from Compound (l) Compound (m):

(wherein TMS is a trimethylsilyl group, and Me is a methyl group.) is produced; and from Compound (m) Compound (n):

(wherein TMS is a trimethylsilyl group, Me is a methyl group, and Et is an ethyl group.) is produced; and from Compound (n) Compound (o):

(wherein TMS is a trimethylsilyl group, Me is a methyl group, and Et is an ethyl group.) is produced; and from Compound (o) Compound (p):

(wherein TMS is a trimethylsilyl group, Me is a methyl group, and Et is an ethyl group.) is produced; and from Compound (p) Compound (q):

(wherein Me is a methyl group, and Et is an ethyl group.) is produced; and from Compound (q) Compound (r):

(wherein Me is a methyl group, Et is an ethyl group, and Pr is a propyl group.) is produced; and from Compound (r) Compound (s):

(wherein Et is an ethyl group, and Pr is a propyl group.) is produced; and from Compound (s) Compound (t):

(wherein Et is an ethyl group, and tBu is a t-butyl group.) is produced; and from Compound (t) Compound (h):

(wherein Et is an ethyl group.) is produced; and wherein it contains one or more steps selected from the group consisting of;

-   (1) a step to obtain Compound (l) mixing Compound (k), alithiating     agent, a formylation reagent and an iodination reagent; -   (2) a step to obtain Compound (m) mixing Compound (l), crotyl     alcohol, triethylsilane and an acid, and reacting said mixture     without use of solvent; -   (3) a step to obtain Compound (l) mixing Compound (v), a by product     in the step (2), with an oxidizing agent and a base; -   (4) a step to obtain Compound (n) mixing Compound (m), a palladium     catalyst, a base and a phase-transfer catalyst, and refluxing said     mixture in solvent; -   (5) a step to obtain Compound (o) mixing Compound (n), an osmium     catalyst, a co-oxidizing agent, a base and an asymmetric reagent; -   (6) a step to obtain Compound (p) mixing Compound (o), a base and     iodine, and refluxing said mixture in an alcohol-water mix liquid; -   (7) a step to obtain Compound (q) mixing Compound (p) and a     desilylation-iodination reagent; -   (8) a step to obtain Compound (r) mixing Compound (q), a palladium     catalyst and a base, and reacting said mixture in 1-propanol under a     carbon monoxide gas atmosphere; -   (9) a step to obtain Compound (s) mixing Compound (r) and a     demethylation reagent, and reacting said mixture at room     temperature; -   (10) a step to obtain Compound (t) reacting Compound (s) under the     presence of t-butyl acrylate and a base.

Further, the invention relates to the above process wherein in the step (1) the lithiating agent is n-butyl lithium.

The invention also relates to the above process wherein in the step (1) the reaction temperature is the constant temperature of −30 to −40° C.

Further, the invention relates to the above process wherein in the step (3) the oxidizing agent is TEMPO-sodium hypochlorite.

The invention also relates to the above process wherein in the step (4) the base is potassium carbonate or N,N-diisopropylethylamine.

Further, the invention relates to the above process wherein in the step (4) the solvent is tetrahydrofuran, or a diisopropyl ether-acetonitrile-water mix liquid.

The invention also relates to the above process wherein in the step (5) the osmium catalyst is potassium osmate (VI).

Further, the invention relates to the above process wherein in step (6) the iodine against Compound (o) is in 4 equivalent.

The invention also relates to the above process wherein in the step (7) the desilylation-iodination reagent is iodine-silver trifluoroacetate or N-chlorosuccinimide-sodium iodide.

Further, the invention relates to the above process, wherein Compound (q) is purified chemically by purification steps comprising a step to add the reaction product obtained by the step to produce Compound (q) from Compound (p) to an aqueous alkaline solution and to stir; a step to add an organic solvent and to stir, followed by removal of the organic layer; and a step to make the aqueous layer acidic and to extract with an organic solvent.

The invention also relates to the above process wherein the aqueous alkaline solution is an aqueous sodium hydoxide solution.

Further, the invention relates to the above process wherein the organic solvent is chloroform.

The invention also relates to the above process, wherein Compound (q) is purified optically by purification steps comprising a step to dissolve the reaction product obtained by the step to produce Compound (q) from Compound (p) in a high polarity solvent, followed by lamination of a low polarity solvent; and a step to filter a precipitate which is followed by concentration of the filtrate to dryness under reduced pressure.

Further, the invention relates to the above process wherein the high polarity solvent is chloroform.

The invention also relates to the above process wherein the low polarity solvent is n-hexane.

Further, the invention relates to the above process wherein in step (10) the base is potassium carbonate.

The invention also relates to use of the tricyclic ketone obtained by the above process to the preparation of camptothecin analogs.

Further, the invention relates to the process for preparing camptothecin analogs wherein the tricyclic ketone obtained by the above process is reacted with 2′-amino-5′-hydroxypropiophenone.

The invention also relates to the above process, wherein the 2′-amino-5′-hydroxypropiophenone is 2′-amino-5′-hydroxypropiophenone obtained by the above process.

Further, the invention relates to the above process, wherein the tricyclic ketone and 2′-amino-5′-hydroxypropiophenone are mixed and said mixture is reacted under an inert atmosphere.

The invention makes it becoming possible to prepare efficiently 2′-amino-5′-hydroxypropiophenone corresponding to the AB-ring part of the CPT skeleton by adopting these constituents and makes it possible to put a total synthesis of CPT into practical use. Additionally, as to the intermediate Compound (c′) and Compound (d′) in the process of the invention there is yet no report of their synthesis, and therefore they are useful novel compounds.

The invention also makes it possible to carry out practically an asymmetric synthesis of compound(s) by adopting these constituents, whereby compound(s) have the skeleton becoming the CDE-ring part (the tricyclic ketone part) in the CPT skeleton.

As to the synthesis of 2′-amino-5′-hydroxypropiophenone of the AB-ring in the CPT skeleton, a process for preparing 2′-amino-5′-hydroxypropiophenone comprises one or more steps of the followings;

-   (1) the step to synthesize 5-benzyloxy-2-nitrobenzaldehyde (Compound     (b′)) from 5-hydroxy-2-nitrobenzaldehyde (Compound (a)); -   (2) the step to synthesize     1-(5-benzyloxy-2-nitrophenol)-2-propen-1-ol (Compound (c′)) from     Compound (b′); -   (3) the step to synthesize     1-(5-benzyloxy-2-nitrophenol)-2-propen-1-one (Compound (d′)) from     Compound (c′); and -   (4) the step to synthesize 2′-amino-5′-hydroxypropiophenone     (Compound (e)) from Compound (d′).

As a typical synthetic route, the following synthetic route:

(wherein R is a protective group which can be deprotected by a catalytic reduction) is shown.

In the invention, in case R is a protective group which can be deprotected by a catalytic reduction, it is not particularly limited, but typical examples are benzyl ether type protective groups such as a benzyl, methoxybenzyl, 2,6-dimethylbenzyl or 4-nitrobenzyl groups, and benzyl carbonate type protective groups such as a benzyloxycarbonyl group, though a benzyl group is expediently used particularly in view of a reagent cost.

Further, as to Compound (a) which is the starting material, that synthesized by a known method, that chemically converted from a similar compound, that isolated and purified from various kinds of natural materials, and natural materials containing Compound (a) can be used. A commercially available reagent may also be used.

In the following the above steps (1) to (4) are explained more specifically.

In the step (1), Compound (a) is dissolved or suspended in solvent, followed by addition of a benzylation reagent and a base and by heating under stirring to afford Compound (b).

As solvent N,N-dimethylformamide (DMF), dimethyl sulfoxide, chloroform, acetonitrile, ethanol, water and the like can be used, and DMF is preferable particularly in view of solubility and reactivity.

The used amount of DMF may be three or more times based on that of Compound (a), preferably in the range of 3 to 20 times.

As a benzylation reagent any one can expediently be used if it is conventionally used. Illustrative of specific examples are benzyl chloride, benzyl bromide, benzyl iodide, phenyldiazomethane, dibenzyl carbonate and the like, and in particular benzyl chloride can expediently be used.

The used amount of a benzylation reagent may appropriately be prepared according to the reagent, though in case of using, for example, benzyl chloride, it is used in 1 to 5 equivalent based on that of Compound (a), preferably 1 to 2 equivalent.

As a base any one can expediently be used if it is conventionally used. Illustrative of specific examples are potassium carbonate, sodium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like, and in particular potassium carbonate can expediently be used.

The used amount of a base may appropriately be prepared according to the reagent, though in case of using, for example, potassium carbonate, it is used in 1 to 10 equivalent based on that of Compound (a), preferably 1 to 4 equivalent.

As a heating temperature it is in the range of 60 to 100° C., preferably 60 to 80° C.

Additionally, the reaction time is in the range of 0.5 to 24 hours, preferably 1 to 20 hours.

In the step (2), Compound (c) is obtained dropping Grignard reagent to Compound (b) under an inert gas atmosphere.

As an inert gas any one may be used in case it is a noble gas such as argon, helium, neon, krypton, xenon, radon or the like, or a gas of low reactivity such as nitrogen, and argon and nitrogen are preferable particularly in view of the cost.

As Grignard reagent any one can expediently be used if it is conventionally used. Illustrative of specific examples are vinyl magnesium bromide, vinyl magnesium chloride, vinyl magnesium iodide and the like, and in particular vinyl magnesium bromide can expediently be used.

The used amount of Grignard reagent may be prepared according to the reagent, though in case of, for example, vinyl magnesium bromide, it is used in 1 to 2 equivalent based on that of Compound (b), preferably 1 to 1.5 equivalent.

In case Grignard reagent is dropped to Compound (b) solution or on the contrary Compound (b) solution is dropped to Grignard reagent, synthesis of Compound (c) is possible, though in order to reduce the production of the reduced type byproduct (hereinafter described as Compound (f))

(wherein R is a protective group which can be deprotected by a catalytic reduction.), it is preferable to drop Grignard reagent to Compound (b) solution.

As a used amount of solvent in the reaction, for example, tetrahydrofuran (hereinafter referred to as THF) may be in an amount of 10 to 100 times, and to reduce particularly production of the alcohol an amount of 50 to 100 times is preferable.

Also, the reaction temperature is preferably not more than 10° C., and to reduce particularly production of the alcohol −78 to −40° C. is preferable.

Additionally, the reaction period is 0.1 to 3 hours, and in particular, preferably, it is 0.5 to 1 hours.

In the step (3), Compound (d) can be obtained mixing Compound (c) with an oxidizing agent and stirring the mixture.

As an oxidizing agent any one can expediently be used if it is conventionally used. Illustrative of such oxidizing agents are, for example, manganese dioxide, Dess-Martin Periodinane, Jones reagent (Na₂Cr₂O₇/H₂SO₄), PCC, PDC, DMSO/oxalyl chloride/triethylamine (Swern oxidation), TEMPO-sodium hypochlorite and the like, and in particular, manganese dioxide, Dess-Martin Periodinane, Jones reagent and TEMPO-sodium hypochlorite can preferably be used.

As to these oxidizing agents, one prepared just before use is preferably used, and in case of, for example, manganese dioxide, one prepared just before use from potassium permanganate and manganese sulfate can expediently be used.

The used amount of an oxidizing agent may appropriately be prepared according to the reagent, though in case of, for example, manganese dioxide, it is used in 2 to 50 times based on that of Compound (c), preferably 4 to 10 times.

As solvent, for example, chloroform, methylene chloride, ethyl acetate, benzene, toluene and the like can expediently be used, and in particular, chloroform and methylene chloride are preferable.

The used amount of solvent is 5 to 50 times, preferably 10 to 20 times.

Further, the reaction time is 1 to 48 hours and in particular, 1 to 18 hours are preferable.

In the step (4), Compound (e) can be obtained by a catalytic reduction of Compound (d).

As a catalyst for reduction palladium-carbon, palladium hydroxide-carbon, rhodium-alumina and the like can expediently be used, and in particular, palladium-carbon and palladium hydroxide-carbon are preferable.

The used amount of a catalyst for reduction is 0.01 to 0.5 equivalent based on that of Compound (d), preferably 0.05 to 0.2 equivalent.

As solvent any one can expediently be used if it is conventionally used, though ethyl acetate is preferable in view of solubility.

The used amount of solvent is 5 to 50 times, preferably 10 to 20 times.

Additionally, the reaction time is 0.1 to 24 hours and in particular, preferably, it is 1 to 3 hours.

Further, instead of synthesizing Compound (e) via the above steps (1) to (4), from Compound (a):

Compound (c″):

is produced; and from Compound (c″) Compound (d″):

is produced; and from Compound (d″) Compound (e):

can be produced. In this synthetic route, Compound (c″) can be obtained dropping Grignard reagent to Compound (a) under an inert atmosphere. Further, Compound (d″) can be obtained mixing Compound (c″) and an oxidizing agent and stirring the mixture, and Compound (e) can be obtained by a catalytic reduction of Compound (d″). Here, Grignard reagent and the oxidizing agent which can be used are the same as those in the above steps (2) and (3). In this synthetic route, since no protecting group is used, the synthesis of the AB-ring part can simply be carried out.

Further, camptothecin analogs can be prepared by reacting Compound (e) obtained in the step (4) or the synthetic route described previously and a tricyclic ketone, though as tricyclic ketone like this, for example, Compound (h):

can be used.

As to synthesis of the CDE-ring part (the tricyclic ketone part) of the CPT skeleton, preparation of the tricyclic ketone is carried out via the following synthetic route.

(wherein TMS is a trimethylsilyl group, Me is a methyl group, Et is an ethyl group, Pr is a propyl group, and ^(t)Bu is a t-butyl group.)

As to the starting Compound (k) in the above synthetic route, that synthesized by the above Curran route (Josien, H,; Ko, S. B.; Bom, D.: Curran, D. P. Chem. Eur. J. 1998, 4 67-83) described previously, that chemically converted from a similar compound that isolated and purified from various kinds of natural materials, or a natural material itself, which contains Compound (k), can be used.

A preferable synthetic process for synthesizing the tricyclic ketone in the above synthetic route contains one or more steps from the 12 steps consisting of;

(1) in the step to synthesize-4-iodo-2-methoxy-6-trimethylsilyl-3-pyridinecarbaldehyde (hereinafter referred to as Compound (1)) from 2-methoxy-6-trimethylsilylpyridine (hereinafter referred to as Compound (k)), n-butyl lithium is used as a base and the reaction is carried out at the constant temperature of to −40° C.;

-   (2) in the step to synthesize     3-(2-butenyloxymethyl)-4-iodo-2-methoxy-6-trimethylsilylpyridine     (hereinafter referred to as Compound (m)) from Compound (l), a     reaction solvent is not used; -   (3) in the step to synthesize Compound (l) from     3-hydroxymethyl-4-iodo-2-methoxy-6-trimethylsilylpyridine     (hereinafter referred to as Compound (v)), TEMPO-sodium hypochlorite     is used as an oxidizing agent; -   (4) in the step to synthesize     4-ethyl-8-methoxy-6-trimethylsilyl-1H-pyrano[3,4-c]pyridine     (hereinafter referred to as Compound (n)) from Compound (m), a mixed     liquid of diisopropylether, acetonitrile and water is used as a     reaction solvent, and N,N-diisopropylethylamine is used as a base; -   (5) in the step to synthesize     (S)-4-ethyl-3,4-dihydro-3,4-dihydroxy-8-methoxy-6-trimethylsilyl-1H-pyrano[3,4-c]pyridine     (hereinafter referred to as Compound (o)) from Compound (n),     potassium osmate(VI) is used as an osmium catalyst; -   (6) in the step to synthesize     (S)-4-ethyl-3,4-dihydro-4-hydroxy-8-methoxy-6-trimethylsilyl-3-oxo-1H-pyrano[3,4-c]pyridine     (hereinafter referred to as Compound (p)) from Compound (o), the     reaction mixture is refluxed using iodine (4 equivalent); -   (7) in the step to synthesize     (S)-4-ethyl-3,4-dihydro-4-hydroxy-6-iodo-8-methoxy-3-oxo-1H-pyrano[3,4-c]pyridine     (hereinafter referred to as Compound (q)) from Compound (p),     N-chlorosuccinimide-sodium iodide is used in acetic acid; -   (8) in the step to purify Compound (q) chemically, the mixture is     added with a basic solution such as aqueous sodium hydroxide     solution to make a solution alkaline, washed with an organic solvent     such as chloroform, and then the water layer after acidification is     extracted with an organic solvent such as chloroform; -   (9) in the step to purify Compound (q) optically, Compound (q) is     dissolved in a high polarity solvent such as chloroform and     laminated with a low polarity solvent such as n-hexane to give     precipitate which is removed by filtration, followed by     concentration of the filtrate; -   (10) in the step to synthesize propyl     (S)-4-ethyl-3,4-dihydro-4-hydroxy-8-methoxy-3-oxo-1H-pyrano[3,4-c]pyridine-6-carboxylate     (hereinafter referred to as Compound (r)) from Compound (q),     palladium acetate is used as a palladium catalyst; -   (11) in the step to synthesize propyl     (S)-4-ethyl-3,4,7,8-tetrahydro-4-hydroxy-3,8-dioxo-1H-pyrano[3,4-c]pyridine-6-carboxylate     (hereinafter referred to as Compound (s)) from Compound (r), the     reaction is carried out at room temperature; -   (12) in the step to synthesize 1,1-dimethylethyl     (S)-4-ethyl-3,4,8,10-tetrahydro-4,6-dihydroxy-3,10-dioxo-1H-pyrano[3,4-f]indolidin-7-carboxylate     (hereinafter referred to as Compound (t)) from Compound (s), Michel     addition is carried out using potassium carbonate.

Further, (13) in the step to obtain SN-38 from (S)-4-ethyl-7,8-dihydro-4-hydroxy-1H-pyrano[3,4-f]indolidin-3,6,10(4H)-trione (hereinafter referred to as Compound (h)) and Compound (e), SN-38 can be expediently be obtained carrying out the reaction in an inert gas atmosphere to afford SN-38 expediently.

In the following, the above 13 steps are explained more specifically.

In the step of (1), Compound (k) is dissolved in solvent, followed by addition of a lithiation, formylation and iodination reagents and stirring to afford Compound (l).

As solvent tetrahydrofuran (THF), diethyl ether, hexane, heptane and the like can be used, and THF is preferable particularly in view of solubility and reactivity.

As a lithiation reagent any one can expediently be used if it is conventionally used. Illustrative of specific examples are n-butyllithium, s-butyllithium, t-butyllithium, lithium diisopropylamide (LDA), lithium bis(trimethylsilyl)amide (LiHMDS) and the like, and n-butyllithium can expediently be used particularly in view of handling and reactivity.

The used amount of a lithiation reagent may appropriately be prepared according to the reagent, though in case of using, for example, n-butyllithium, it is used in 2 to 10 equivalent based on that of Compound (k), preferably 2 to 5 equivalent.

Illustrative of specific examples of a formylation reagent are N-formyl-N,N′,N′-trimethylethylenediamine, dimethylformamide (DMF) and the like, and N-formyl-N,N′,N′-trimethylethylenediamine is expediently used considering the subsequent iodination.

The used amount of a formylation reagent, for example, in case of using N-formyl-N,N′,N′-trimethylethylenediamine is used in 1 to 10 equivalent based on that of Compound (k), preferably 1 to 3 equivalent.

As an iodination reagent iodine, N-iodosuccinimide (NIS) and the like can be used, and iodine is preferable particularly in view of the cost and reactivity.

The used amount of an iodination reagent is used in 1 to 10 equivalent based on that of Compound (k), preferably 1 to 5 equivalent.

The reaction temperature is in the range of 0 to −78° C., preferably the constant temperature of −30 to −40° C.

In the step of (2), Compound (l) is added with crotyl alcohol, triethylsilane and an acid and stirred without using solvent to afford Compound (m).

As the used amount of crotyl alcohol, it is used in 1 to 10 equivalent based on that of Compound (k), preferably 2 to 5 equivalent.

As the used amount of triethylsilane, it is used in 1 to 10 equivalent based on that of Compound (k), preferably 1 to 4 equivalent.

As an acid trifluoroacetic acid (TFA), sulfuric acid, methanesulfonic acid, hydrochloric acid and the like can be used, and TFA is preferable particularly in view of reactivity.

The used amount of an acid, for example, in case of TFA is 1 to 15 equivalent based on that of Compound (l), preferably 5 to 10 equivalent.

In the step of (3), Compound (l) can be obtained by dissolving Compound (v), a byproduct in the step of (2), insolvent, followed by addition of an oxidizing agent and a base, and stirring.

As solvent any one can expediently be used if it is conventionally used. Illustrative of such solvent are dichloromethane, chloroform, acetonitrile, toluene, n-hexane and the like, and toluene and n-hexane are preferable particularly in view of reactivity.

Illustrative of oxidizing agents are manganese dioxide, Dess-Martin Periodinane, Jones reagent (Na₂Cr₂O₇/H₂SO₄), PCC, PDC, DMSO-oxalyl chloride-triethylamine (Swern oxidation), TEMPO-hypochlorite and the like, and in particular, TEMPO-hypochlorite is preferable, more preferably TEMPO-sodium hypochlorite.

As the used amount of an oxidizing agent, for example, in case of TEMPO-sodium hypochlorite, TEMPO is used in 0.001 to 0.1 equivalent based on that of Compound (v), preferably 0.005 to 0.02 equivalent. Additionally, sodium hypochlorite is used in 1 to 5 equivalent, preferably 1 to 2 equivalent.

As a base any one can expediently be used if it is conventionally used. Illustrative of such bases are sodium bicarbonate, sodium carbonate, potassium carbonate, calcium carbonate, sodium hydroxide, calcium hydroxide, triethylamine and the like, with sodium bicarbonate being particularly preferable.

As the used amount of a base, for example, in case of sodium bicarbonate, it is used in 1 to 10 equivalent based on that of Compound (v), preferably 2 to 4 equivalent.

The reaction temperature is in the range of −10 to 30° C., preferably not −10 to 10° C. particularly to suppress a side reaction.

Additionally, the reaction period is in the range of 0.5 to 10 hours, preferably 0.5 to 5 hours.

In the step of (4), Compound (m) is dissolved in solvent, added with a palladium catalyst, a base and a phase-transfer catalyst, and refluxed to afford Compound (n).

As solvent acetonitrile, tetrahydrofuran (THF), diisopropyl ether (IPE), diethyl ether, toluene, water and the like can be used, and acetonitrile, THF, IPE and water are preferable particularly in view of reactivity, more preferably THF or an acetonitrile-IPE-water mix liquid.

As a palladium catalyst palladium acetate, tetrakis-(triphenylphosphine)palladium, dichlorobis-(triphenylphosphine) palladium, palladium chloride and the like can expediently be used, and palladium acetate is preferable particularly in view of reactivity.

The used amount of a palladium catalyst is in 0.01 to 1 equivalent based on that of Compound (m), preferably 0.05 to 0.2 equivalent.

As a base any one can expediently be used if it is conventionally used. Illustrative of such bases are, for example, sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), sodium hydroxide, potassium hydroxide and the like, with potassium carbonate and DIPEA being particularly preferable.

The used amount of a base, for example, in case of DIPEA is in 1 to 20 equivalent based on that of Compound (m), preferably 5 to 10 equivalent.

As a phase-transfer catalyst any one can expediently be used if it is a quaternary ammonium salt or crown ether which are conventionally used, with tetrabutylammonium bromide being particularly preferable.

The used amount of a phase-transfer catalyst, for example, in case of tetrabutylammonium bromide is in 0.1 to 3 equivalent based on that of Compound (m), preferably 0.5 to 1.5 equivalent.

Further, the reaction period in case of using THF is in the range of 1 to 20 hours, preferably 4 to 10 hours. In case of using an acetonitrile-IPE-water mix liquid it is in the range of 0.5 to 10 hours, preferably 1 to 5 hours.

In the step of (5), Compound (n) is dissolved in an alcohol-water mix liquid, added with an osmium catalyst, a co-oxidizing agent, an asymmetric catalyst, a base and methanesulfonamide, and stirred to afford Compound (o).

Illustrative of alcohols are methanol, ethanol, 1-propanol, isopropanol (IPA), 1-butanol, 2-butanol, t-butyl alcohol and the like, and t-butyl alcohol is preferable particularly in view of reactivity.

As an osmium catalyst osmium tetraoxide, potassium osmate (VI) and the like can expediently be used, and potassium osmate(VI) is preferable particularly in view of handling.

The used amount of an osmium catalyst is in 0.001 to 0.1 equivalent based on that of Compound (n), preferably 0.002 to 0.01 equivalent.

As a co-oxidizing agent potassium hexacyanoferrate(III), N-methylmorpholine N-oxide (NMO) and the like can expediently be used, and potassium hexacyanoferrate(III) is preferable particularly in view of the reactivity.

The used amount of a co-oxidizing agent, for example, in case of potassium hexacyanoferrate(III), is used in 1 to 10 equivalent based on that of Compound (n), preferably 2 to 5 equivalent.

Illustrative of asymmetric catalysts are (DHQD)₂PYR, (DHQD)₂PHAL, (DHQD)₂AQN and the like, and (DHQD)₂PYR is preferable particularly in view of the optical yield.

The used amount of an asymmetric catalyst, for example, in case of (DHQD)₂PYR, is used in 0.005 to 0.1 equivalent based on that of Compound (n), preferably 0.01 to 0.05 equivalent.

As a base sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like can be used, and potassium carbonate is preferable particularly in view of the reactivity.

The used amount of a base, for example, in case of potassium carbonate, is in 1 to 20 equivalent based on that of Compound (n), preferably 4 to 10 equivalent.

The used amount of methanesulfonamide is in 0.1 to 5 equivalent based on that of Compound (n), preferably 0.5 to 2 equivalent.

The reaction temperature is in the range of −10 to 30° C., preferably −10 to 10° C.

In the step of (6), Compound (o) is dissolved in solvent, added with a base and iodine, and refluxed to afford Compound (p).

Illustrative of solvent are methanol, ethanol, 1-propanol, isopropanol (IPA), water and the like, with a methanol-water mix liquid being particularly preferable in view of reactivity.

As a base any one can expediently be used if it is conventionally used. Illustrative of such bases are sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like, with calcium carbonate being particularly preferable.

The used amount of a base, for example, in case of calcium carbonate, is in 1 to 10 equivalent based on that of Compound (o), preferably 2 to 5 equivalent.

The used amount of iodine is in 1 to 10 equivalent based on that of Compound (o), preferably 3 to 5 equivalent. Additionally, the reaction period is in the range of 0.5 to 20 hours, preferably 1 to 5 hours.

In the step of (7), Compound (p) is dissolved in solvent, and reacted under iodine-silver trifluoroacetate (hereinafter referred to as I₂-CF₃COOAg) or N-chlorosuccinimide-sodium iodide (hereinafter referred to as NCS-NaI) to afford Compound (q).

As to solvent, in case of I₂-CF₃COOAg dichloromethane, carbon tetrachloride, chloroform and the like are expedient, and in particular, dichloromethane is preferable. Additionally, in case of NCS-NaI acetic acid, acetonitrile and the like can be used, and acetic acid is preferable particularly in view of reactivity.

As to the used amount of I₂-CF₃COOAg, I₂ is used in 1 to 10 equivalent based on that of Compound (p), preferably 2 to 4 equivalent. Additionally, CF₃COOAg is used in 1 to 10 equivalent, preferably 2 to 4 equivalent.

As to the used amount of NCS-NaI, NCS is used in 1 to 20 equivalent based on that of Compound (p), preferably 5 to 8 equivalent. Additionally, NaI is used in 1 to 20 equivalent based on that of Compound (p), preferably 5 to 8 equivalent.

The reaction temperature in case of using I₂-CF₃COOAg is 10 to 60° C., preferably 20 to 40° C. Further, in case of using NCS-NaI it is 20° C. to a reflux temperature, preferably 50 to 80° C.

Additionally, the reaction period is in the range of 5 to 48 hours, preferably 15 to 24 hours.

In the step of (8), Compound (q) is added with a basic solvent, for example, such as aqueous 0.2N sodium hydroxide, and stirred to give the lactone-ring opening compound (Compound (u)):

(wherein Me is a methyl group, and Et is an ethyl group.), which is soluble in the aqueous basic solution. When washing the solution with an organic solvent, a neutral-basic substance moves to an organic layer. The organic layer is separated, followed by acidification of the water layer with an acid and extraction with an organic solvent to recover Compound (q) in good purity.

The basic solvent is in the range of 0.01 to 5 N, preferably 0.1 to 1 N, more preferably 0.2 to 0.5 N.

Illustrative of used bases are potassium hydroxide, calcium hydroxide, sodium hydroxide, potassium carbonate, sodium carbonate and the like, with sodium hydroxide being particularly preferable.

As an organic solvent any one can expediently be used if it is conventionally used. Illustrative of such solvent are dichloromethane, chloroform, ethyl acetate, toluene, diethyl ether, diisopropyl ether and the like, and in particular, with dichloromethane and chloroform being particularly preferable.

Illustrative of used acids are hydrochloric acid, sulfuric acid, nitric acid, acetic acid, phosphoric acid, trifluoroacetic acid and the like, with hydrochloric acid being particularly preferable.

In the step of (9), Compound (q) is dissolved in a high polarity solvent, and laminated with a low polarity solvent to precipitate crystals which are filtered. The filtrate is concentrated under reduced pressure to dryness. The obtained crystals are racemic, and a more optically purified Compound (q) is obtained as a residue.

As a high polarity solvent chloroform, dichloromethane, ethyl acetate, methanol, ethanol, propanol and the like can be used, and in particular, chloroform is preferable.

The used amount of a high polarity solvent, for example, in case of chloroform, is 1 to 10 ml, preferably 3 to 6 ml, against Compound (q) 1 g.

Illustrative of low polarity solvent are n-hexane, n-heptane, diethyl ether and the like, with n-hexane being particularly preferable.

The ratio of a high polarity solvent: a low polarity solvent is, for example, in case of chloroform:n-hexane, is in the range of 10:1 to 1:20, preferably 2:1 to 1:2.

The temperature in the crystallization procedure is not more than room temperature, preferably not more than 5° C.

In the step of (10), Compound (q) is dissolved in 1-propanol, added with a palladium catalyst and a base, and reacted under a carbon monoxide gas atmosphere to afford Compound (r).

As to a palladium catalyst, palladium acetate, tetrakis-(triphenylphosphine)palladium, dichlorobis-(triphenylphosphine)palladium, palladium chloride and the like can expediently be used, and palladium acetate is preferable particularly in view of the reactivity.

The used amount of a palladium catalyst is in 0.005 to 0.5 equivalent based on that of Compound (q), preferably 0.01 to 0.1 equivalent.

As a base any one can expediently be used if it is conventionally used. Illustrative of such bases are, for example, sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), sodium hydroxide, potassium hydroxide and the like, with potassium carbonate being particularly preferable.

The used amount of a base, for example, in case of potassium carbonate, is in 1 to 20 equivalent based on that of Compound (q), preferably 4 to 10 equivalent.

The reaction temperature is in the range of 20° C. to a reflux temperature, preferably not more than 50° C. to a reflux temperature.

In the step of (11), Compound (r) is dissolved in solvent, added with a demethylation reagent, and reacted at room temperature to afford Compound (s).

As solvent acetonitrile, chloroform, dichloromethane, toluene and the like can be used, and in particular, acetonitrile is preferable.

Illustrative of demethylation reagents are chlorotrimethylsilane-sodium iodide, iodotrimethylsilane, hydriodic acid, hydrobromic acid and the like, with chlorotrimethylsilane-sodium iodide being particularly preferable.

The used amount of a demethylation reagent, for example, in case of chlorotrimethylsilane-sodium iodide, is in 1 to 10 equivalent based on that of Compound (r), preferably 2 to 5 equivalent.

In the step of (12), Compound (s) is dissolved in solvent, added with a base, and stirred under an inert gas atmosphere. The obtained mixture is dropped with t-butyl acrylate, and stirred under an inert gas atmosphere to afford Compound (t)

As to solvent, dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF) and the like can expediently be used, and DMSO is preferable particularly in view of reactivity.

As a base potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide and the like can be used, and in particular, potassium carbonate is preferable.

The used amount of a base, for example, in case of potassium carbonate, is in 1 to 20 equivalent based on that of Compound (s), preferably 2 to 5 equivalent.

As an inert gas any one may be used in case it is a noble gas such as argon, helium, neon, krypton, xenon, radon or the like, or a gas of low receptivity such as nitrogen, and argon and nitrogen are preferable particularly in view of the cost.

The used amount of t-butyl acrylate is in 1 to 20 equivalent based on that of Compound (s), preferably 8 to 12 equivalent.

The reaction temperature is in the range of 20 to 80° C., preferably 40 to 60° C.

Further, the reaction period is in the range of 5 to 48 hours, and in particular, it is preferably not more than 24 hours in order to avoid particularly decomposition of Compound (t) produced.

In the step of (13), Compound (h) and Compound (e) are dissolved in solvent, added with an acid, and heated under an inert gas atmosphere and stirring to afford SN-38.

As solvent toluene, acetic acid and the like can expediently be used, and in particular, a toluene-acetic acid mix liquid is preferable.

As an inert gas any one may be used in case it is a noble gas such as argon, helium, neon, krypton, xenon, radon or the like, or a gas of low reactivity such as nitrogen, and argon and nitrogen are preferable particularly in view of the cost.

As an acid toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid and the like can be used, and toluenesulfonic acid is preferable particularly in view of reactivity.

The used amount of an acid, for example, in case of toluenesulfonic acid, is 1 to 100 mg based on that of Compound (h) 1 g, preferably 10 to 30 mg.

The used amount of Compound (e) is in 1 to 3 equivalent based on that of Compound (h), preferably 1 to 1.5 equivalent.

The reaction temperature is in the range of 50° C. to a reflux temperature, preferably 80° C. to a reflux temperature.

In the following, the invention will be illustrated in more detail by way of examples, but the invention is not limited to these.

EXAMPLE 1 Synthesis of Compound (b′)

Wherein Bn is a benzyl group.

Compound (a) (38.5 g, 0.230 mol) was dissolved in 116 mL of DMF or acetone. Potassium carbonate (33.4 g, 0.242 mol, 2.1 eq.) and 27.8 mL (0.242 mol, 1.05 eq.) or 59.95 mL (0.461 mol, 2 eq.) of benzyl chloride were added to the stirred solution of Compound (a) at room temperature under argon atmosphere. After the addition, the mixture was heated at 60° C. and vigorously stirred for 20 hours with periodical checks the content of Compound (a). After Compound (a) was not detected anymore, the mixture was filtered by suction.

The solid material was washed with the same solvent used for the reaction. The filtrate and the washing were combined, and the solvent was evaporated under reduced pressure. And water (300 mL) was added to the residue. The mixture was stirred and the precipitates were filtered by suction and dried in air. After air-drying, the filtered material was dissolved in 170 mL of ethyl acetate. This solution was added to 1 L of hexane with stirring. The precipitated solid material was filtered by suction, washed with 300 mL of a mixture of ethyl acetate and hexane (1:10) and dried under reduced pressure.

Experiment (Exp.) 1 and 2, in which were discriminated by the amount of the benzyl chloride was varied and in Exp. 3, acetone was used as the reaction solvent.

TABLE 1 Reaction Amount of benzyl Reaction Isolated solvent chloride time yield Exp. 1 DMF 1.05 eq. 20 hours 94% Exp. 2 DMF 2.00 eq.  1 hour 94% Exp. 3 acetone 2.00 eq. 18 hours — —: less than limit of detection

As shown in Table 1, 20 hours were required for the completion of the reaction with the yield of 94% when 1.05 eq. of benzyl chloride was used (Exp. 1). When 2.00 eq. of the benzyl chloride was used (Exp. 2), the reaction was finished in 1 hour and the yield was 94%. Three-fold amounts of DMF was minimal requirement for the reaction, otherwise the stirring was disturbed by precipitating the solid material during the reaction. When acetone was used as the reaction solvent, the reaction did not proceed even under reflux condition for 18 hours.

HPLC Operation Conditions

-   -   Column: Inertsil ODS-2, 5 μm, 4.6 mm ID×250 mm (GL science-made)     -   Temperature: constant temperature about 40° C.     -   Mobile phase: water:acetonitrile mixture (1:1)     -   Flow rate: 1 mL/min     -   Detect: 220 nm

EXAMPLE 2 Synthesis of Compound (c′) (1)

Wherein Bn is a benzyl group. Compound (b′) (1.0 g, 3.89 mmol) was dissolved in 20 mL of THF. Vinyl magnesium bromide (1.0 M THF solution, 5.84 mL, 5.84 mmol, 1.5 eq.) was added dropwise to the ice-cooled, stirred solution of Compound (b′) under argon atmosphere over 15 minutes. During the addition, the internal temperature was kept within 3 to 10° C. After the stirring for 1 hour, the reaction solution was added to a saturated aqueous solution of ammonium chloride (20 mL) with stirring, and then to the solution 20 mL of ethyl acetate and 4 mL of hexane were added, and the obtained organic layer was washed with 20 mL of water and an aqueous saturated solution of sodium chloride successively, and dried over 3 g of sodium sulfate. The solvent was evaporated under reduced pressure to give Reaction product A.

A THF solution (20 mL) of Compound (b′) prepared as mentioned above was added dropwise to an ice-cooled solution of vinyl magnesium bromide (1.0 M THF solution, 5.84 mL) under argon atmosphere over 15 minutes. During the addition, the internal temperature was kept within 3 to 10° C. After the stirring for 1 hour, the reaction solution was added to a saturated aqueous solution of ammonium chloride (20 mL) with stirring and then to the solution 20 mL of ethyl acetate and 4 mL of hexane were added, and the organic layer was separated and washed with 20 mL of water and a saturated aqueous solution of sodium chloride and dried over 3 g of sodium sulfate. The solvent was evaporated under reduced pressure to give Reaction product B.

Reaction product A and B were purified through silica gel column chromatography (ethyl acetate:hexane=1:20) to give Exp. 4 from Reaction product A and Exp. 5 from Reaction product B, respectively.

TABLE 2 Yield of Compound (f) Yield of Compound (c′) (peak area %) Exp. 4 84.0% 3.5% Exp. 5 26.8% 11.3%

As shown in Table 2, when the Grignard reagent was added to the solution of Compound (b′), the yield of the product increased by 57% and formation of Compound (f), a byproduct, was suppressed.

HPLC operation conditions; refer to the Example 1.

EXAMPLE 3 Synthesis of Compound (c′) (2)

Compound (b′) (1.0 g, 3.89 mmol) was dissolved in 10 to 100 mL of THF. Vinylmagnesium bromide (1.0 M in THF, 5.84 mL, 5.84 mmol, 1.5 eq.) was added dropwise to the stirred solution of Compound (b′) under argon atmosphere over 15 minutes. After the stirring for 1 hour, the reaction solution was added to a saturated aqueous solution of ammonium chloride (20 mL) with stirring and then to the solution 20 mL of ethyl acetate and 4 mL of hexane were added, and the organic layer was separated and washed with 20 mL of water and a saturated aqueous solution of sodium chloride successively, and dried over 3 g of sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by the same manner described in Example 2 (Exp. 4 and 5). Exp. 6 represents the results of the reaction at 20° C. using 20-fold amount of the solvent. Exp. 7 to 9 represent the results of the reaction at 3° C. using 10-fold, 40-fold, 100-fold amount of the solvent, respectively. The results of the reactions are summarized in Table 3.

TABLE 3 Yield of Yield of Reaction Amount of Compound (c′) Compound (f) temperature solvent (peak area %) (peak area %) Exp. 4 3° C. 20-fold 84.0% 3.5% Exp. 6 20° C.  20-fold 68.7% 4.8% Exp. 7 3° C. 10-fold 81.1% 5.7% Exp. 8 3° C. 40-fold 88.6% 3.5% Exp. 9 3° C. 100-fold  90.2% 2.8%

As shown in Table 3, when the reaction was carried out at 10° C. or lower, more preferred 5° C. or lower, the formation of Compound (f) was suppressed and the yield of Compound (C′) increased by 15% or more. When 100-fold amount of the solvent was used (Exp. 9), the formation of Compound (f) was suppressed and the yield of Compound (c′) increased by 6%.

HPLC operation conditions; refer to Example 1.

EXAMPLE 4 Synthesis of Compound (d′) (1)

Wherein Bn is a benzyl group. (1) Preparation of Manganese Dioxide:

An aqueous solution of manganese sulfate penta-hydrate (122 g/150 mL, 0.506 mol) and 117 mL of 40% sodium hydroxide were added to an aqueous solution of potassium permanganate (96.0 g/600 mL, 0.607 mol) at room temperature with stirring. After stirring for 18 hours, the solid material was filtered by suction and washed with water. The obtained solid material was dried in air to give 91.2 g of manganese dioxide.

(2) Synthesis of Compound (d′)

Compound (c′) (2.00 g, 7.02 mmol) was dissolved in 20 mL of chloroform, dichloromethane or ethyl acetate. Manganese dioxide 8.00 g (4-fold amount, 92.0 mmol, 13 eq.) prepared by the above mentioned method was added to the vigorous stirred solution of Compound (c′) at 25° C. under argon atmosphere. The mixture was vigorously stirred for 15 hours. After the starting material was not detected anymore, the mixture was filtered by suction. The obtained solid material was washed with 20 mL of chloroform. The filtrate and the washing were combined, and the solvent was evaporated under reduced pressure. Exp. 10 to 12 were obtained.

TABLE 4 Remained starting Reaction material Reaction solvent time (peak area %) Yield Exp. 10 chloroform 15 hours — 91% Exp. 11 dichloromethane  3 hours — 79% Exp. 12 ethyl acetate 24 hours 8% — —: less than limit of detection

As shown in Table 4, when chloroform or dichloromethane was used as the reaction solvent, Compound (d′) was synthesized in good yields. In particular, the reaction time was shortened to one third when dichloromethane was used as the reaction solvent. On the other hand, Compound (c′) was remained even after 24 hours when ethyl acetate was used.

EXAMPLE 5 Synthesis of Compound (d′) (2)

An aqueous solution of sodium hypochlorite (available chlorine min. 5.0%; 42 mL) and sodium hydrogen carbonate aqueous solution (7.1 g in 60 mL of water) were added to a vigorously stirred, ice-cooled mixture of 7.0 g (3.5 mmol) of Compound (c′), toluene (70 mL), ethyl acetate (70 mL), water (10 mL) and 38.3 mg (1 mol %) of TEMPO under ice cooling (at 2 to 6° C., 55 min). After 5 minutes, 0.4% (HPLC, peak area %) of the starting material was detected. The mixture was placed still and the separated organic layer taken and was washed with a mixture of potassium iodide and potassium hydrogen sulfate (yellow→red-brown), a saturated aqueous solution of sodium thiosulfate and then water, successively. The solvent was evaporated under reduced pressure to give 6.4 g of Compound (d′) (yield 91%, purity 92.6% by HPLC), which was purified by recrystallization from a mixture of methanol and water (25:1) to give 2.3 g of purified Compound (d′) (starting from 3.0 g; purified Compound (d′), 2.3 g, recovery: 77%, purity: 95.2% by HPLC).

HPLC operation conditions; refer to Example 1.

EXAMPLE 6 Synthesis of Compound (e)

Wherein Bn is a benzyl group.

To an ice-cooled stirred solution of 1.84 g (6.50 mmol) of Compound (d′) in 37 mL of ethyl acetate, 0.69 g (0.65 mmol, 10 mol %) of 10% palladium carbon was added under argon atmosphere. The mixture was vigorously stirred at 25° C. under hydrogen atmosphere and a part of the mixture was taken as a sample for HPLC, periodically. The reaction mixture was filtered and the filtrate was evaporated. Exp. 13 to 14 were obtained.

TABLE 5 Yield of Yield of Compound Compound (g) Reaction time (e) (peak area %) Exp. 13 0.1 hour   71% 14% Exp. 14 13 hours 81%  0%

As shown in Table 5, the reaction was conducted for longer than 13 hours, the yield of Compound (e) increased by 10% and formation of the by-product, Compound (g), was suppressed.

-   -   HPLC Operation Conditions     -   Column: Inertsil ODS-2, 5 μm, 4.6 mm ID×250 mm (GL science-made)     -   Temperature: constant temperature about 40° C.     -   Mobile phase: water:acetonitrile mixture (1:1)     -   Flow rate: 1 mL/min     -   Detect: 254 nm

EXAMPLE 7 Whole Synthetic Process of 2′-amino-5′-hydroxypropiophenone

The whole synthetic process of 2′-amino-5′-hydroxypropiophenone is as follows.

(1) Synthesis of Compound (b′)

Compound (a) (1.00 g, 5.98 mmol) was dissolved in 3 mL of DMF. Potassium carbonate (0.87 g, 6.28 mmol, 2.1 eq.) and 0.72 mL (6.28 mmol, 1.05 eq.) of benzyl chloride were added to the stirred solution of Compound (a) at room temperature under argon atmosphere. After the addition, the mixture was heated at 60° C. and vigorously stirred for 20 hours with periodical checks confirmation of the content of Compound (a) by HPLC. After Compound (a) was not detected anymore, the mixture was filtered by suction.

The solid material was washed with 3 mL of DMF. The filtrate and the washing were combined, and the solvent was evaporated under reduced pressure. After the evaporation, the residue was added to 100 mL of water. After the mixture was stirred for a while, the insoluble material was filtered by suction and dried in air. After the air-drying, the material was dried under reduced pressure (1 mmHg, at 20° C.) to give 1.45 g (yield 95%) of Compound (b′) as pale yellow solid. The physical properties of Compound (b′), including NMR spectrum, are as follows. Compound (b′); mp 71-73° C.

¹H-NMR (400 MHz, CDCl₃): δ 5.21 (2H, s, PhCH₂O), 7.21 (1H, dd, J=2.8, 9.3 Hz), 7.35-7.44 (6H, m), 8.16 (1H, d, J=9.3 Hz), 10.48 (1H, s, CHO).

IR (KBr): 1250, 1333, 1514, 1589, 1697 cm⁻¹.

EI-MS: m/z 257 (M⁺).

(2) Synthesis of Compound (c′)

Compound (b′) (1.0 g, 3.89 mmol) was dissolved in 20 mL of THF. A solution of vinylmagnesium bromide (1.0 M in THF solution, 5.84 mL, 5.84 mmol, 1.5 eq.) was added dropwise over 15 minutes to the ice-cooled stirred solution of Compound (b′) under argon atmosphere. During the addition, the internal temperature was kept within 3 to 10° C. After the stirring for 1 hour, the reaction solution was added to a stirred saturated aqueous solution ammonium chloride. Then 20 mL of ethyl acetate and 4 mL of hexane were added, and the organic layer was separated and washed with 20 mL of water and a saturated aqueous solution of sodium chloride and dried over 3 g of sodium sulfate. The solvent was evaporated under reduced pressure. The residue (1.19 g) was purified through silica gel column chromatography (ethyl acetate:hexane=1:20) to give 0.93 g of Compound (c′) (yield 84%) as orange solid. The physical properties of Compound (c′), including NMR spectrum, are as follows. Compound (c′); mp 60-63° C.

¹H-NMR (400 MHz, CDCl₃) δ: 5.15 (2H, s, PhCH₂O), 5.22-5.26 (1H, m), 5.39-5.44 (1H, m), 5.90 (1H, d, J=5.1 Hz), 6.06 (1H, ddd, J=5.1, 10.5, 15.6 Hz), 6.94 (1H, dd, J=2.9, 9.0 Hz), 7.34 (1H, d, J=2.9 Hz), 7.35-7.44 (5H, m), 8.04 (1H, d, J=9.0 Hz).

IR (KBr): 3298, 1614, 1582, 1506, 1292, 1229 cm⁻¹.

EI-MS: m/z 285 (M⁺).

(3) Synthesis of Compound (d′)

Compound (c′) (2.00 g, 7.02 mmol) was dissolved in 20 mL of chloroform. Manganese dioxide (8.00 g, 4-fold amount, 92.0 mol, 13 eq.) was added to the vigorously stirred solution of Compound (c′) at 25° C. under argon atmosphere. The mixture was vigorously stirred for 15 hours. After the starting material was not detected anymore, the mixture was filtered by suction. The obtained solid material was washed with 20 mL of chloroform. The filtrate and the washing were combined, and the solvent was evaporated under reduced pressure. The residue was purified through silica gel column chromatography (ethyl acetate:hexane=1:20) to give 1.88 g of Compound (d′) (yield 95%) as white solid. The physical properties of Compound (d′) including NMR spectrum are as follows.

Compound (d′); mp 84-85° C.

¹H-NMR (400 MHz, CDCl₃) δ: 5.17 (2H, s, PhCH₂O), 5.83 (1H, d, J=17.7 Hz), 6.01 (1H, d, J=10.6 Hz), 6.62 (1H, dd, J=10.6, 17.7 Hz), 6.91 (1H, d, J=2.7 Hz), 7.10 (1H, dd, J=2.7, 9.0 Hz), 7.37-7.43 (5H, m), 8.17 (1H, d, J=9.0 Hz).

IR (KBr): 1686, 1578, 1506, 1342, 1244 cm⁻¹.

EI-MS: m/z 283 (M⁺)

(4) Synthesis of Compound (e)

To an ice-cooled stirred solution of 1.84 g (6.50 mmol) of Compound (d′) in 37 mL of ethyl acetate, 0.69 g (0.65 mmol, 10 mol %) of 10% palladium carbon was added under argon atmosphere. The mixture was vigorously stirred at 25° C. under hydrogen atmosphere. After stirring for 13 hours, the catalyst was removed by filtration from reaction mixture. The filtrate was evaporated under reduced pressure to give 0.87 g (yield 81%, purity 91.14% by HPLC) of the crude product as orange solid. 500 mg of the obtained reaction product was purified through silica gel column chromatography (ethyl acetate:hexane=1:10→1:4) to give 421 mg of Compound (e) (yield 84%, purity 95.59% by HPLC) as yellow solid. The physical properties of Compound (e) including NMR spectrum are as follows.

Compound (e); mp 131-140° C.

¹H-NMR (400 MHz, CDCl₃) δ: 1.20 (3H, t, J=7.2 Hz), 2.93 (2H, q, J=7.2 Hz), 6.59 (1H, d, J=8.8 Hz), 6.88 (1H, dd, J=2.9, 8.8 Hz), 7.23 (1H, d, J=2.9 Hz).

IR (KBr): 3379, 3296, 1670, 1447, 1194 cm⁻¹.

EI-MS: m/z 165 (M⁺).

EXAMPLE 8 Synthetic Method of Compound (e) without Protective Group R

(1) Synthesis of Compound (c″) from Compound (a)

Compound (a) (500 mg, 2.99 mmol) was dissolved in 15 mL of THF. To the ice-cooled stirred solution of Compound (a), vinylmagnesium bromide (1.0 M in THF, 7.5 mL, 7.5 mmol, 2.5 eq.) was added dropwise over about 5 minutes under argon atmosphere. After the stirring for 1 hour, the reaction mixture was added to ice-cooled 1 mol/L hydrochloric acid (30 mL). Then 30 mL of ethyl acetate and 5 mL of hexane were added, and the organic layer was separated and washed with 50 mL of water and a saturated aqueous solution of sodium chloride and dried over 3 g of sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified through silica gel column chromatography (ethyl acetate:hexane=1:10→1:3) to give 541 mg of Compound (c″) (yield 93%) as yellow-brown solid.

Compound (c″);

¹H-NMR (400 MHz, CDCl₃) δ: 5.22-5.26 (1H, m), 5.35-5.40 (1H, m), 5.90-5.92 (1H, m), 6.06 (1H, ddd, J=5.2, 10.5, 15.6 Hz), 6.83 (1H, dd, J=2.7, 9.0 Hz), 7.19 (1H, d, J=2.7 Hz), 8.00 (1H, d, J=9.0 Hz).

(2) Synthesis of Compound (d″) from Compound (c″)

Compound (c″) (1.00 g, 5.13 mmol) was dissolved in 8 mL of acetone. To the ice-cooled solution of Compound (c″), Jones reagent (3.0 mL, 5 mmol, 1.5 eq.) was added with stirring. After the stirring for 0.5 hours, three pieces of ice and a saturated aqueous solution of sodium hydrogen sulfate (5 mL) were added to the reaction mixture. Then 50 mL of ethyl acetate and 5 mL of hexane were added, and the layer was separately and washed with 50 mL of water and a saturated aqueous solution of sodium chloride successively and dried over 5 g of sodium sulfate. The solvent was evaporated under reduced pressure to give 0.82 g of Compound (d″) (yield 83%).

Compound (d″);

¹H-NMR (400 MHz, DMSO-d₆) δ: 5.84 (1H, d, J=17.6 Hz), 6.11 (1H, d, J=10.7 Hz), 6.60 (1H, dd, J=10.7, 17.7 Hz), 6.75 (1H, d, 2.7 Hz), 7.03 (1H, dd, 9.1 Hz), 8.13 (1H, d, J=9.1 Hz), 11.41 (1H, s).

(3) Synthesis of Compound (e) from Compound (d″)

Compound (d″) (100 mg, 0.513 mmol) was dissolved in 1 mL of ethyl acetate. To the ice-cooled solution 55 mg (0.0513 mmol, 10 mol %) of 10% palladium carbon was added under argon atmosphere with stirring. The mixture was stirred at room temperature under hydrogen atmosphere for 18 hours. The catalyst was filtrated and the filtrate was evaporated under reduced pressure to give 64 mg of Compound (e) (yield 76%) as yellow solid.

EXAMPLE 9 Synthesis of 7-ethyl-10-hydroxycamptothecin (SN-38)

Compound (e) (0.36 g, 2.14 mmol) obtained in Example 7 and Compound (h) (0.50 g, 1.82 mmol) were suspended in a mixture of acetic acid and toluene (AcOH-toluene; 1:1, 10 mL). p-Toluenesulfonic acid monohydrate (p-TsOH.H₂O; 10 mg) was added to the suspension at room temperature, and the mixture was stirred at 100° C. for 18 hours. The reaction mixture was condensed under reduced pressure, toluene (10 mL) was added to the residue, and mixture was condensed under reduced pressure again. Acetone (9 mL) was added to the residue and the mixture was stirred at room temperature 2 hours, the insoluble material was filtered and washed with acetone (2 mL, twice). The filtered material was dried under reduced pressure to give SN-38 (0.63 g, purity 97.7% by HPLC, yield 89%) as black solid.

HPLC operation conditions

-   -   Column: Inertsil ODS-2, 5 μm, 4.6 mm ID×250 mm (GL science-made)     -   Temperature: constant temperature about 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: methanol-acetonitrile-10 mM potassium         dihydrogenphosphate (1:1:3)     -   Detect: 254 nm         SN-38;

¹H-NMR (400 MHz, CDCl₃) δ: 0.98 (3H, t, J=7 Hz, CH₃), 1.38 (3H, t, J=7 Hz, CH₃), 1.90 (2H, q, J=7 Hz, CH₂), 3.08 (2H, q, J=7 Hz, CH₂), 5.17 (2H, s, CH₂O), 5.23 (1H, d, J=16 Hz), 5.54 (1H, d, J=16 Hz), 6.83 (1H, d, J=9 Hz), 7.34-7.39 (3H, m).

EXAMPLE 10 Synthesis of 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (SN-38B-11)

SN-38B-11 (1.22 g, 2.08 mmol, yield 89%, enantiopurity 99.8% ee) was obtained from SN-38 (0.91 g, 2.32 mmol) synthesized in Example 9 by the reported procedure (Sawada, S.; Okajima, S.; Aiyama, R.; Nokata, K.; Furuta, T.; Yokokura, T.; Sugino, E.; Yamaguchi, K.; Miyasaka, T. Chem. Pharm. Bull. 1991, 39, 1446.).

-   -   Chiral HPLC Operation Conditions     -   Column: DAICEL CHIRALCEL OD-H, 0.46 cmID×25 cm (#ODHOCE-AK031)     -   Guard cartridge: DAICEL CHIRALCEL OD-H, 0.4 cmID×1 cm     -   Injection amount: 10 μg/10 μL     -   Temperature: constant temperature about 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: dimethylamine:hexane:ethanol mixture (1:250:250)     -   Detect: 254 nm

EXAMPLE 11 Synthesis of 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11)

SN-38B-11 (1.00 g, 1.7 mmol) obtained in Example 10 was suspended in 1/10 N hydrochloric acid (20 mL, 2.0 mmol), and the suspension was heated at about 80° C. to dissolve in. Acetonitrile (100 mL) was added to the solution, and the mixture was stirred at room temperature for overnight. The precipitates were filtered, dried, and humidified under 75% RH afforded CPT-11 (0.95 mg, yield 89.8%) as pale yellow crystalline powder.

EXAMPLE 12 Synthesis of Compound (l) (1)

Compound (l) was obtained by formylation of Compound (k) at around −30° C. or −20° C. with n-butyl lithium and N-formyl-N,N′,N′-trimethylethylenediamine following iodination at around −30° C. or −20° C. with n-butyl lithium and iodine.

Compound (k) (5.0 g; 0.028 mol) was dissolved in anhydrous THF (about 66 mL) under nitrogen gas atmosphere. The mixture was cooled at around −30° C. or −20° C. n-Butyl lithium (1.6 mol/L in hexane; 21.2 mL, 0.034 mol, 1.2 eq.) was added dropwise to the solution and the mixture was stirred under cooling. Then N-formyl-N,N′,N′-trimethylethylenediamine (4.4 g, 0.0034 mol, 1.2 eq.) was added to the reaction mixture as the formylation reagent and the mixture was at stirred under cooling.

n-Butyl lithium (1.6 mol/L in hexane; 35 mL, 0.05 mol, 1.2 eq.) was added dropwise to the mixture and stirred at the temperature shown in Table 6. Then iodine (18.4 g) in anhydrous THF (19 mL) was added dropwise to the stirred mixture.

An aqueous solution of sodium hydrogen sulfite (12 g in 200 mL) was added to the mixture. After stirring, the recovered organic layer (hexane) was analyzed by HPLC. The results were shown in Table 6.

-   -   HPLC Operation Conditions     -   Column: Capcell Pack ODS UG120, 4.6 mmID×150 mm     -   Mobile phase: 50 mM potassium dihydrogenphosphate acetonitrile         mixture (9:11)     -   Detect: 220 nm     -   Flow rate: about 1 mL/min     -   Temperature: room temperature

TABLE 6 Reaction Reaction Folmylation time Iodination time Compound Compound Yield (° C.)¹⁾ (hour) (° C.)¹⁾ (hour) (k)³⁾ (l)³⁾ (%)⁴⁾ Exp. −48 to −30 3.0 −70 to −65 0.3 NT⁵⁾ 67.8 70.6 15 −32 to −29 around −75 Exp. −35 to −28 1.0 −30 to −20 0.5 5.9 67.8 71.9 16 around −35 −35 to −25 Exp. −20 to −15 2.0 −10 to −5 0.5 6.2 70.5 66.7 17 −20 to −15 −10 to −5 Exp. −10 to −5 3.0 −10 to 0 0.5 3.4 77.6 63.7 18 −10 to −5 −10 to 0 ¹⁾The upper lines show the actual internal temperature during the addition. The lower lines show the actual internal temperature range during the stirring. ²⁾Exp. 15: the results of an experiment under the reported conditions ³⁾Peak area % ⁴⁾The yields were corrected by purity (HPLC; peak area %) ⁵⁾NT: Not tested

As shown in Table 6, Compound (l) was obtained in 60% yield or more when n-butyl lithium was used as the lithiation reagent.

As shown in Exp. 16, the reaction was proved to proceed at constant temperature at −40° C. to −30° C. in good yield (more than 70%).

EXAMPLE 13 Purification of Compound (l) (Washing with Diluted Hydrochloric Acid)

Compound (k) (5.0 g, 0.028 mol) was dissolved in anhydrous THF (about 66 mL). The reaction was carried out as described in Example 12 at constant temperature at around −35° C. The n-hexane layer obtained from the reaction mixture was washed with diluted hydrochloric acid (the same amount of the organic layer).

After the washing, the organic layer was dried over sodium sulfate, filtered and a part of the filtrate was analyzed by HPLC under the condition given in Example 12. The results were shown in Table 7.

TABLE 7 The analytical results of each the hexane layer after washed with diluted hydrochloric acid hexane Com- Com- layer¹⁾ HCl Residue pound pound Recovery (mL) (mol/L) (g) (k)²⁾ MTPC^(2,3)) (l)²⁾ (%) 25 —⁴⁾ 1.5 6.0 11.8 54.7 — 50 0.1 2.9 6.6 12.4 58.7 100 50 1.0 2.6 1.8 13.0 61.2 100 50 2.5 2.6 0.4 12.6 62.4 100 50 3.5 2.6 0.2 12.7 64.2 100 ¹⁾The hexane layer obtained from the reaction mixture was divided into 5 parts (25 mL for the intact mixture and 4 parts of 50 mL for washing). Each the 50 mL part was washed with diluted HCl listed in Table 7 and the recovered hexane layers were analyzed by HPLC. ²⁾peak area % ³⁾MTPC; 2-methoxy-6-trimethylsilylpyridine 3-carbaldehyde ⁴⁾not washed

As shown in Table 7, Compound (k) was almost removed by washing with the diluted hydrochloric acid. When 1.0 mol/mL and more concentrated hydrochloric acid was used for the washing, Compound (l) was obtained in good purity. 2-methoxy-6-trimethylsilylpyridine 3-carbaldehyde (MTPC), the formylated intermediate of Compound (k), was hardly removed by this method.

EXAMPLE 14 Purification of Compound (l) (Stepwise Washings with Diluted Hydrochloric Acid)

Compound (k) (5.0 g, 0.028 mol) was dissolved in anhydrous THF (about 66 mL). The reaction was carried out as described in Example 12 at constant temperature at around −35° C. The n-hexane layer obtained from the reaction mixture was washed in turn (top to bottom) with the diluted hydrochloric acid listed in Table 8 (the same amount of the organic layer).

After the washing, the aqueous acidic layer was separated, neutralized with sodium carbonate and then extracted with n-hexane. The organic layer was dried over magnesium sulfate, filtered and a part of the filtrate was analyzed by HPLC under the conditions given in Example 12. The results are summarized in Table 8.

TABLE 8 The analytical results on the hexane extracts from the neutralized aqueous layer, which was obtained by washing the original hexane layer with diluted hydrochloric acid in order as follows (top to bottom). Residue Compound Compound Recovery HCl (mol/L) (g)¹⁾ (k)²⁾ (l)²⁾ (%) Washing — NT³⁾ NT — with water 0.1 0.40 24.9 10.7 — 0.1 0.04 NT NT — 1.0 0.21 67.0 13.7 — 2.5 0.28 71.0  3.0 — 5.0 0.54 13.0  4.0 — Residue⁴⁾ 7.27 ND⁵⁾ 77.9 98.3 ¹⁾Each the diluted HCl washing was neutralized with sodium carbonate and the mixture was extracted with n-hexane. The organic layer was dried, filtered and the filtrate was evaporated under reduced pressure to dryness. ²⁾HPLC (peak area %) ³⁾Not tested ⁴⁾The residue of the hexane layer after the stepwise washings. ⁵⁾Not detected: less than limit of detection

-   -   As shown in Table 8, the hexane layer was conveniently purified         by multi step washings with hydrochloric acid of different         concentrations to give Compound (l) with high purity.

EXAMPLE 15 Purification of Compound (l) (Purification by Distillation)

Compound (k) (5.0 g, 0.028 mol) was dissolved in anhydrous THF (about 66 mL). The reaction was carried out as described in Example 12 at constant temperature at around −35° C. The obtained reaction mixture (n-hexane layer) was recovered and distilled at 81 to 99° C. under reduced pressure (around 0.35 mmHg). After distillation, the residue in the distillation vessel was purified through silica gel column chromatography with n-hexane, and then a mixture of n-hexane and ethyl acetate (50:1) to give the purified product.

The residue and the purified material were analyzed by HPLC under the conditions below. The results are shown in the table 9.

-   -   HPLC Operation Conditions     -   Column: Capcell Pack ODS UG120, 4.6 mmID×150 mm     -   Mobile phase: 50 mM potassium dihydrogenphosphate:acetonitrile         mixture (1:1)     -   Wave length: 220 nm     -   Flow rate: about 1 mL/min     -   Temperature: room temperature

TABLE 9 The analytical results on the fractions of the residue of the hexane layer by distillation Compound Compound Recovery (g) (k)¹⁾ MTPC¹⁾ (l)¹⁾ (%) Intact — 3.6 13.5 71.5 — mixture Fraction-1 0.19 47.3 36.1 8.7 — Fraction-2 1.16 8.9 53.7 28.8 — Trap 1.17 70.3 ND²⁾ ND — Residue 5.13 0.3 3.1 89.9 75.9 Purified — — 3.9 95 product³⁾ ¹⁾peak area % ²⁾Not detected: less than detection limit ³⁾The final residue was purified through silica gel column chromatography

As shown in Table 9, MTPC was almost removed by distillation. Further purification by silica gel column chromatography afforded Compound (l) with excellent purity. It is not preferred that the distillation at higher temperature than that in Table 9 because the coloration and decomposition of Compound (l) were observed.

EXAMPLE 16 Purification of Compound (l) (Recovery as hydrochloric acid salt)

Compound (k) (5.0 g, 0.028 mol) was dissolved in anhydrous THF (about 66 mL). The reaction was carried out as described in Example 12 at constant temperature around −35° C. The reaction mixture (10 g) was dissolved in 10 N hydrochloric acid (10 mL) and stirred at room temperature. The yellow precipitates were filtered and washed with a small amount of 10 N hydrochloric acid and the material was dissolved in water (about 10 mL). The pH of the aqueous solution was adjusted to about 8 by adding sodium hydrogen carbonate, and the mixture was extracted with hexane and the organic layer was evaporated under reduced pressure to dryness.

The residue was analyzed by HPLC under the conditions given in Example 15. The results are shown in Table 10.

TABLE 10 The analytical results on the extracts by neutralizing the hydrochloric acid salt obtained in 10 mol/L hydrochloric acid. (g)¹⁾ MTP²⁾ MTPC²⁾ Compound (l)²⁾ Recovery (%) Pre- 10 —³⁾ 16.9 61.8 — purified Post- 6 — 3.3 90.0 87.4 purified ¹⁾the weight of the residue ²⁾peak area % ³⁾Not tested

As shown in Table 10, the crude reaction product was collected as the hydrochloric acid salt and the salt was recovered as Compound (l) by neutralization, MTPC was almost removed by this method.

Compound (l): yellow oil.

¹H-NMR (499 MHz, CDCl³) δ: 0.30 (9H, s) 4.05 (3H, s), 7.67 (1H, s), 10.19 (1H, s),

EI:MS:m/z 335 (M⁺).

EXAMPLE 17 Synthesis of Compound (m)

To a mixture of Compound (l) (20.0 g, 56.0 mmol, content: 93.9% by HPLC), triethylsilane, (17.9 mL, 112.0 mmol, 2 eq.) and crotyl alcohol (15.7 mL, 184.8 mmol, 3.3 eq.), trifluoroacetic acid (28.5 mL, 375.3 mmol, 6.7 eq.) was added dropwise at 0 to 5° C. under nitrogen gas atmosphere with stirring. After stirring at the temperature for 30 min, the mixture was stirred at ambient temperature for 20 hrs. An aqueous solution of sodium carbonate (20.8 g in 277 mL of water) and n-hexane (56 mL) were added to the mixture and the organic layer was separated and the aqueous layer was extracted with n-hexane (56 mL). The combined organic layers were evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography; silica gel (80 g, Fuji Silysia PSQ100B) with a mixture of n-hexane-ethyl acetate (73:3) as the eluent.

The results of this Example is summarized Table 11 (Exp. 20). Exp. 19 in the table shows the results of a trace experiment under the reported conditions; Josien, H.; Ko, S. B.; Bom, D.; Curran, D. P., Chem. Eur. J. 1998, 4, 67-83. Curran, D. P.; Ko, S. B.; Josien, H., Angew. Chem. Int. Ed. Engl. 1995, 34, 2683-2684. Under the reported conditions, dichloromethane was used as the reaction solvent. The equal level of the product (m) was obtained in the quality and yield without dichloromethane.

TABLE 11 HPLC (% peak area) Solvent Time (h) (l) (m) (v) others Exp. 19 CH₂Cl₂ 17 1.19 68.08 16.94 13.79 Exp. 20 neat 20 0.40 64.38 24.40 10.82 HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: Constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: acetonitrile-10 mM potassium dihydrogenphosphate         (5:1)     -   Detect: 254 nm

EXAMPLE 18 Synthesis of Compound (l) (2)

To a mixture of Compound (v) (1.00 g, content; 98.43%, 2.9 mmol), TEMPO (2.3 mg, 0.015 mmol, 0.005 eq.) and 7% (w/v) sodium hydrogen carbonate (7.0 mL) in toluene (8.7 mL), an aqueous solution of sodium hypochlorite (available chlorine; minimal 5%, 4.5 g, 3.0 mmol, 1.05 eq.) was added at 0 to 5° C. and then the mixture was stirred at 0 to 5° C. for 2 hrs. 10% sodium sulfite (3.7 mL, 2.9 mmol) was added to the mixture and the resulting mixture was stirred at 0 to 5° C. for 30 min. The insoluble material in the mixture was removed by filtration and the material on the filter paper was washed with toluene (1 mL×3). The organic layer of the filtrate was separated and washed with water (10 mL), dried over sodium sulfate (2 g), filtered and the desiccant was washed with toluene. The filtrate and the washing were combined and then evaporated under reduced pressure to dryness. Compound (l):yellow oil, 0.93 g (87% yield), content: 90.60% by HPLC (see Example 17).

EXAMPLE 19 Synthesis of Compound (n) (1)

Compound (m) (1.60 g) was dissolved in the solvents listed in Table 12, tetrabutylammonium bromide (0.83 g), potassium carbonate (0.71 g) and palladium acetate (57 mg) were added to the solution. Each the reaction was conducted under the conditions directed in Table 12.

The reaction mixture was poured into ice-cooled n-hexane (18 mL) with stirring. The insoluble material was filtered by suction and the material was washed with n-hexane (6 mL×3). The filtrate and the washing were combined and washed with water (9 mL×2), dried over anhydrous sodium sulfate and then evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography with n-hexane-ethyl acetate (95:5) as the eluent.

Exp. 21 in Table 12 was the results of an experiment conducted under the reported conditions; Josien, H.; Ko, S. B.; Bom, D.; Curran, D. P., Chem. Eur. J. 1998, 4, 67-83. Curran, D. P.; Ko, S. B.; Josien, H., Angew. Chem. Int. Ed. Engl. 1995, 34, 2683-2684.

The ratio of the endo and exo forms of each the purified product was measured by HPLC. As shown in Table 12, satisfactory selectivity (endo-exo ratio) and isolated yield were obtained when THF was used as the reaction solvent under refluxed conditions (Exp. 25-27). The isolated yields of (Exp. 25-27) were higher than that of the reported conditions (Exp. 21) by 10% or more.

TABLE 12 Yield Solvent Temperature Time (h) Ratio (%) Exp. 21 DMF 85° C. 1.5 2.3 69 Exp. 22 CHCl₃ Reflux 5.0 3.1 — Exp. 23 Tol 85° C. 96.0 3.7 — Exp. 24 MeCN 85° C. 2.0 4.3 — Exp. 25 THF Reflux 4.0 6.6 79 Exp. 26 THF Reflux 5.0 7.0 84 Exp. 27 THF Reflux 4.0 7.1 82 DMF: N,N-dimethylformamide, Tol: toluene, MeCN: acetonitrile, THF: tetrahydrofuran, Ratio: peak area of endo form/peak area of exo form (HPLC), Yield: isolated yield, —: not determined, HPLC operating conditions; see Example 17.

EXAMPLE 20 Synthesis of Compound (n) (2)

To a solution of Compound (m) (1.27 g, 2.6 mmol, content; 78.7%) in a mixture of diisopropyl ether-acetonitrile-water (4:3:1, 20 mL), tetrabutylammonium bromide (0.82 g, 2.6 mmol), N,N-diisopropylethylamine (3.48 mL, 20.8 mmol, 8 eq.) and palladium acetate (57 mg, 0.26 mmol) were added and the resulting mixture was heated under reflux for 30 min. After cooling the inner temperature to 20° C. or below, the insoluble material in the mixture was filtered by suction and the filtered material was washed with n-hexane (2.6 mL×3). To the combined filtrate and the washing, n-hexane (10 mL) and 10% sodium sulfite (16 mL, 13.0 mmol, 5 eq.) were added. The organic layer was separated and washed with 1 N hydrochloric acid (16.4 mL) and then water (10 mL×2), continuously. The organic solution was evaporated under reduced pressure to dryness. Compound (n); tanned oil, 0.83 g (91% yield), content: 73.3% by HPLC (see Example 17), endo-exo ratio: 10.6.

Thus, the selectivity was markedly improved as compared with that of Exp. 21 in Table 12. The isolated yield was also improved by 20% of that of Exp. 21 in Table 12.

EXAMPLE 21 Synthesis of Compound (o)

To a solution of potassium ferricyanide (195.7 g, 0.59 mol), potassium carbonate, (82.1 g, 0.59 mol) and methanesulfonamide (37.7 g, 0.40 mol) in water (990 mL), (DHQD)₂PYR (4.36 g, 4.95 mmol) and potassium osmate (VI) dihydrate (1.0 mmol) were added and the mixture was stirred at around 5° C. for 1 hr. To the stirred mixture, Compound (n) (77.8 g, 0.18 mol, content: 61.5%) was added and the resulting mixture was stirred at the temperature for additional 20 hrs. Powdered sodium sulfite (74.9 g) was added to the mixture and the stirring was continued at the temperature for 30 min. The insoluble material in the mixture was filtered on a Celite pad and the material on the pad was washed with ethyl acetate (4 times, total 770 mL). The organic layer of the filtrate was separated and the aqueous layer was further extracted with ethyl acetate (770 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography with a mixture of dichloromethane-ethyl acetate (4:1) as the eluent; silica gel (280 g, Fuji Silysia PSQ100B). Compound (o); umber solid.

As shown in Table 13, using potassium osmate as the oxidant effected the equal level of the isolated yield and enantioexcess in place of highly volatile osmium (VIII) oxide.

TABLE 13 Oxidant Yield (%) % ee Exp. 28 OsO₄ 82 to 95 95.6 to 96.2 Exp. 29 K₂OsO₄•2H₂O 94 95.9

Exp. 28: conducted under the reported conditions; Josien, H.; Ko, S. B.; Bom, D.; Curran, D. P., Chem. Eur. J. 1998, 4, 67-83. Curran, D. P.; Ko, S. B.; Josien, H., Angew. Chem. Int. Ed. Engl. 1995, 34, 2683-2684.

% ee: Compound (o) obtained here was converted into Compound (p) by the method described in Example 22 and its enantioexcess was measured by chiral HPLC method (see; Example 22)

EXAMPLE 22 Synthesis of Compound (p)

To a solution of Compound (o) (70 g) in a mixture of in ethanol and water (10:1, 1.0 L), iodine (the amount is given in Table 14) and powdered calcium carbonate (47.1 g) were added at ambient temperature with stirring. The reaction mixture was stirred at given temperature and period in Table 14.

One litter of 10% sodium sulfite and chloroform (1.0 L) were added to the mixture and the resulting mixture was stirred at ambient temperature for 30 min. The insoluble material was removed by filtration and the organic layer of the filtrate was separated. The aqueous layer was extracted with chloroform (500 mL×2). The extracts were combined, dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness.

Using 4 equivalent of iodine under reflux conditions, this reaction was completed in 5 hrs, about one tenth of that of the reported conditions with comparable yields.

TABLE 14 Iodine Time (eq.) Temperature (hr) Yield (%) Exp. 30 9 Ambient 48 86 Exp. 31 4 Ambient 72 86 Exp. 32 4 40° C. 48 88 Exp. 33 4 60° C. 20 88 Exp. 34 4 Reflux 5 84 Exp. 30: conducted under the reported conditions;

Josien, H.; Ko, S. B.; Bom, D.; Curran, D. P., Chem. Eur. J. 1998, 4, 67-83.

Curran, D. P.; Ko, S. B.; Josien, H., Angew. Chem Int. Ed. Engl., 1995, 34, 2683-2684.

Yield (%): isolated yields

HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: 10 mM potassium dihydrogenphosphate-acetonitorile         (4:3)     -   Detect: 254 nm         Chiral HPLC Operating Conditions     -   Column: DAICEL CHIRALCEL OD-H, #ODHOCE-AK031, 0.46 cm ID×25 cm     -   Guard cartridge: DAICEL CHIRALCEL OD-H, 0.4 cm ID×1 cm     -   Temperature: constant temperature at around 25° C.     -   Flow rate: 0.5 mL/min     -   Mobile phase: a mixture of n-hexane-ethanol (200:1)     -   Detect: 254 nm

EXAMPLE 23 Synthesis of Compound (q)

To a solution of Compound (p) (50.2 g) in the solvent (about 400 mL; given in Table 15), the reagents in the table were added and the resulting mixture was stirred at the given temperature for the hours. To the reaction mixture 20% sodium carbonate (1.7 L), 10% sodium sulfite (1.0 L) and chloroform (550 mL) were added with stirring. The organic layer was separated and the aqueous layer was extracted with chloroform (550 mL×2). The extracts were combined, dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. The content of Compound (q) in the residue was quantified by HPLC. The results are summarized in Table 15.

-   -   This conversion was satisfactorily performed by using NCS-NaI at         65° C. in acetic acid (Exp. 39). The period required for the         completion was apparently shortened and the yields of         Compound (q) under the conditions were higher than that of the         report [Comparative Experiment 1 (Com. 1)] by 50% or more.

TABLE 15 Time Yield Solvent Reagent Eq. Temperature (hr) (%) Com. 1 *) ICI 4 RT to 40° C. 48 45 Exp. 35 AcOH NIS 12 65° C. 45 63 Exp. 36 CH₂Cl₂ I₂ - 2 Ambient 17 97 CF₃CO₂Ag Exp. 37 AcOH NCS - NaI 6 65° C. 16 95 Exp. 38 AcOH NCS - NaI 6 65° C. 16 93 Exp. 39 AcOH NCS - NaI 6 65° C. 15 94 *) a mixture of dichloromethane and chloroform (3:2), AcOH: acetic acid, ICl: iodine monochloride, NIS: N-iodosuccinimide, NCS: N-chlorosuccinimide, Eq.: molar ratio of the reagent(s) employed, Yield: isolated yields.

EXAMPLE 24 Purification of Compound (q) (1)

Compound (q) (63 g, Purity 89.2% by HPLC) obtained in Example 23 was suspended in methanol (150 mL), to the suspension was added dropwise 0.2 N sodium hydroxide with vigorous stirring and the stirring was continued for 2 hrs. The alkaline solution was washed with chloroform (400 mL×3) and the pH of the aqueous layer was adjusted with 6 N hydrochloric acid to 1-2 and the acidified solution was extracted with chloroform (400 mL×3). The chloroform layer was dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. Compound (q) (Exp. 40); purity 97.7% (% peak area; HPLC operating conditions: see Example 25)

EXAMPLE 25 Purification of Compound (q) (2)

Compound (q) (50 g) purified by the method described in Example 24 was dissolved in chloroform (240 mL) and n-hexane (400 mL) was softly added on the surface of the solution. The mixture was left to stand at ambient temperature for 15 hrs. The precipitates of the mixture were removed by filtration and the filtrate was evaporated under reduced pressure to dryness (Exp. 41).

Compound (q) (Exp. 40) obtained in Example 24 (93-96% enantioexcess) was optically purified by this method. Compound (q) (Exp. 41) obtained here exhibited 99.7-99.9% enantioexcess by the chiral HPLC given below.

HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: a mixture of acetonitrile-10 mM potassium         dihydrogenphosphate (5:3)     -   Detect: 254 nm         Chiral HPLC Operating Conditions     -   Column: DAICEL CHIRALPAK AD-H, # ADH0CE-BC037, 0.46 cm ID×25 cm     -   Guard cartridge: DAICEL CHIRALPAK AD-H, 0.4 cm ID×1 cm     -   Temperature: constant temperature at around 25° C.     -   Flow rate: 1 mL/min     -   Mobile phase: a mixture of n-hexane-2-propanol (25:1)     -   Detect: 254 nm

EXAMPLE 26 Synthesis of Compound (r)

A solution of Compound (q) (42.8 g, 0.10 mol, content: 84.5%), palladium acetate (1.34 g, 6.0 mmol) and potassium carbonate (24.7 g, 0.18 mol) in propanol (490 mL) was charged in a reaction vessel. The vessel was degassed by suction and released with nitrogen gas, and degassed again by suction and then replaced with carbon monoxide. The mixture was stirred at 60° C. under carbon monoxide atmosphere for 18 hrs. The insoluble material was filtered on a Celite pad and the material was washed with ethyl acetate (300 mL). To the filtrate, 1 N hydrochloric acid (150 mL) and brine (300 mL) were added and the mixture was shaken vigorously. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography; silica gel (100 g, Fuji Silysia PSQ100B) with a mixture of chloroform and methanol (99:1) as the eluent. Compound (r): umber oil, 30.3 g (70% yield), content: 73.4% quantified by HPLC.

HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: 10 mM potassium dihydrogenphosphate-acetonitrile         (4:3)     -   Detect: 254 nm

EXAMPLE 27 Synthesis of Compound (s)

To a stirred solution of Compound (r) (28.7 g, 68.2 mmol, content: 73.4%) and sodium iodide (27.6 g, 0.18 mol) in absolute acetonitrile (141 mL), chlorotrimethylsilane (23.3 mL, 0.18 mmol) was added dropwise at ambient temperature under nitrogen gas atmosphere. The mixture was stirred at ambient temperature for 3 hrs. To the mixture, 1 N hydrochloric acid (8 mL) and 10% sodium sulfite (232 mL) were added and the resulting mixture was stirred for 30 min. The mixture was extracted with ethyl acetate and the organic layer was separated and evaporated under reduced pressure to dryness. Compound (s): 22.3 g (95% yield), content: 85.6% quantified by HPLC (as follows).

HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: 10 mM potassium dihydrogen phosphate-acetonitrile         (5:2).     -   Detect: 254 nm

EXAMPLE 28 Synthesis of Compound (t)

A solution of Compound (s) (0.50 g) in dimethylsulfoxide (DMSO, 7 mL) in the presence of an inorganic base (potassium or cesium carbonate, 0.4 g) was stirred at 50° C. under argon atmosphere for 20 min. To the stirred mixture tert-butyl acrylate (1.8 g) was added dropwise and the resulting mixture was stirred at 50° C. under argon atmosphere for 24 hrs. Water (10 mL) and concentrated hydrochloric acid (1 mL) were added to the ice-cooled mixture and the mixture was extracted with a mixture of toluene and ethyl acetate (4:1, 7 mL×4). The combined extracts were dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to dryness (Exp. 42-43). The residue was assayed by HPLC (as follows).

As shown in Table 16, Compound (t) was obtained in 72% yield using cesium carbonate as the base (Exp. 42), on the other hand, inexpensive potassium carbonate was used as the base, the yield was equal level with the Exp. 42.

TABLE 16 Base Yield (%) Exp. 42 Cs₂CO₃ 72 Exp. 43 K₂CO₃ 77 HPLC Operating Conditions

-   -   Column: GL Science Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: 10 mM potassium dihydrogenphosphate-acetonitrile         (5:2)     -   Detect: 254 nm

EXAMPLE 29 Synthesis of SN-38

A mixture of Compound (h) (0.50 g, 1.82 mmol, content: 96.6%) and Compound (e) (0.36 g, 2.18 mmol) in a mixture of toluene-acetic acid (1:1, 10 mL) in the presence of p-TsOH.H₂O (10 mg) was heated at 90° C. with stirring under nitrogen gas atmosphere for 7 hrs. After cooling, the mixture was evaporated under reduced pressure to dryness. After 2 times of azeotropic removal of acetic acid with toluene (10 mL), acetone (9 mL) was added to the residue and the suspension was stirred under nitrogen atmosphere for 30 min. The solid was collected by filtration, washed with acetone (2 mL×2) and then dried under reduced pressure. SN-38 (Exp. 45): ocherous solid, 0.63 g (89.1% yield), purity; 99.6% by HPLC (see Example 9).

Exp. 44 in Table 17 shows the results of an experiment under the reported conditions; Henegar, K. E.; Ashford, S. W.; Baughman, T. A.; Sih, J. C.; Gu, R. L., J. Org. Chem. 1997, 62, 6588-6597.

Under nitrogen gas atmosphere, the purity and yield of SN-38 are improved as shown in Table 17.

TABLE 17 Purity (%) Yield (%) Exp. 44 Open vessel 97.6 75 Exp. 45 N₂ atm. 99.6 89

EXAMPLE 30 Synthesis of Tricyclic Ketone

Whole synthetic processes of tricyclic ketone (h) starting from Compound (l) are given below;

(1) Synthesis of Compound (m)

To a stirred mixture of Compound (l) (20.0 g, 56.0 mmol, 2 eq., content: 93.9%), triethylsilane (17.9 mL, 112 mmol, 2 eq.) and crotyl alcohol (15.7 mL, 184.8 mmol, 3.3 eq.), trifluoroacetic acid (28.5 mL, 375.2 mmol, 6.7 eq.) was added dropwise at 0-5° C. under nitrogen atmosphere and the mixture was stirred at the temperature for 30 min. The mixture was stirred at ambient temperature for 20 hrs and then an aqueous solution of sodium carbonate (20.8 g in 277 mL of water) and n-hexane (56 mL) were added to the mixture. The organic layer was separated and the aqueous layer was extracted with n-hexane (57 mL). The combined organic layers were evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography; silica gel (80 g, Fuji Silysia PSQ100B), eluent; n-hexane-ethyl acetate (73:3) to remove byproduct, Compound (v) (4.95 g, 14.68 mmol, purity 98.43%, yield 26%). Compound (m); 17.8 g (64% yield), content: 80.0% by HPLC (see Example 17).

¹H-NMR (400 MHz, CDCl₃) δ: 0.24 (9H, s, TMS), 1.69 (3H, dd, J=1.0, 6.1 Hz, ═CHCH₃ ), 3.85-4.05 (2H, m, OCH₂ CH═), 3.93 (3H, s, CH₃O), 4.55 (2H, s, OCH₂), 5.55-5.83 (2H, m, CH═CH), 7.4 7 (1H, s).

Compound (v); 5.0 g (26% yield), content: 98.4% (HPLC).

¹H-NMR (400 MHz, CDCl₃) δ: 0.27 (9H, s, TMS), 2.45 (1H, t, J=6.8 Hz, OH), 3.99 (3H, s, CH₃O), 4.79 (2H, d, J=6.8 Hz, CH₂ OH), 7.49 (1H, s).

(2) Synthesis of Compound (n)

A mixture of Compound (m) (1.27 g, 2.56 mmol, content: 78.73%), tetrabutylammonium bromide (0.82 g, 2.56 mmol) and palladium acetate (57 mg, 0.26 mmol) in a mixture of diisopropyl ether-acetonitrile-water (4:3:1, 20 mL) was heated under reflux for 30 min. After cooling the inner temperature to 20° C. or below, the insoluble material of the mixture was removed by filtration and the material was washed with n-hexane (10 mL). The filtrate and the washing were combined and to the solution, n-hexane (10 mL) and 10% sodium sulfite (16 mL, 113 mmol, 5 eq.) were added. The organic layer of the mixture was separated and washed with 1 N hydrochloric acid (16.4 mL) and water (10 mL×2), successively. The organic layer was evaporated under reduced pressure to dryness. Compound (n) brown oil, 0.83 g (91% yield), content: 73.34% by HPLC, endo-exo ratio: 10.6 by HPLC (see Example 17).

¹H-NMR (400 MHz, CDCl₃) δ: 0.26 (9H, s, TMS), 1.12 (3H, t, J=7.3 Hz, CH₂CH₃ ), 2.31 (2H, dq, J=1.0, 7.3 Hz, CH₂ CH₃), 3.94 (3H, s, OCH₃), 5.00 (2H, s, OCH₂), 6.51 (1H, t, J=1.0 Hz, OCH═), 6.83 (1H, s, aromatic-H).

(3) Synthesis of Compound (o)

To a solution of potassium ferricyanide (195.7 g, 0.59 mol), potassium carbonate (82.1 g, 0.59 mol) and methanesulfonamide (37.7 g, 0.40 mol) in water (990 mL), (DHQD)₂ PYR (4.36 g, 4.95 mmol) and potassium osmate (VI) dihydrate (0.99 mmol) were added and the mixture was stirred at around 5° C. for 1 hr. To the mixture, Compound (n) (77.8 g, 0.18 mol, content: 61.5%) was added and the resulting mixture was stirred at around 5° C. for 20 hrs and then powdered sodium sulfite (74.9 g) was added. The suspension was stirred at around 5° C. for 30 min and the insoluble material was filtered on a Celite pad. The material on the pad was washed with ethyl acetate (4 times, total 770 mL). The organic layer of the filtrate was separated and the aqueous layer was further extracted with ethyl acetate (770 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography; silica gel (700 g, Fuji Silysia PSQ100B), eluent: dichloromethane-ethyl acetate (4:1). Compound (o): umber solid.

(4) Synthesis of Compound (p)

A mixture of Compound (o) (70.2 g), iodine (183.7 g, 0.72 mol) and calcium carbonate (36.23 g, 0.36 mol) in methanol-water (10:1, 1.0 L) was heated under reflux for 5 hrs. After cooling, 10% sodium sulfite (1.0 L) and chloroform (1.0 L) were added to the mixture and the resulting mixture was stirred at ambient temperature for 15 min. The insoluble material was filtered by suction and the material was washed with chloroform (0.5 L). The combined organic layers of the filtrate and the washing were separated and the aqueous layer was further extracted with chloroform (0.5 L). The combined organic layers were dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to dryness. Compound (p) umber oil, 53.6 g (overall 81% yield from Compound (m)), content: 80.4% by HPLC (see Example 22), 96.2% ee by chiral HPLC (see Example 22).

¹H-NMR (400 MHz, CDCl₃) δ: 0.28 (9H, s, TMS), 0.94 (3H, t, J=7.4 Hz, CH₂CH₃ ), 1.76 (2H, q, J=7.4 Hz, CH₂ CH₃), 3.61 (1H, s, OH), 3.98 (3H, s, OCH₃), 5.23 (1H, d, J=15.6 Hz,), 5.54 (1H, d, J=15.6 Hz), 7.33 (1H, s, aromatic-H).

(5) Synthesis of Compound (q)

A mixture of Compound (p) (50.2 g, 0.14 mol, content: 80.4%, 96.2% ee), N-chlorosuccinimide (107.36 g, 0.80 mol) and sodium iodide (120.52 g, 0.80 mol) in acetic acid (411 mL) was warmed at about 65° C. with stirring for 16 hrs. After cooling, 20% sodium carbonate (1.7 L), 10% sodium sulfite (1.0 L) and chloroform (0.6 L) were added, successively, to the mixture. The organic layer of the mixture was separated and the aqueous layer was extracted with chloroform (0.6 L×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and then evaporated under reduced pressure to dryness (crude q).

(6) Purification of Compound (q) (1)

A suspension of crude Compound (q) (the residue of the previous step, purity: 89.2% by HPLC) in methanol (150 mL) was added dropwise to 0.2 N sodium hydroxide (0.40 mol) with stirring. The mixture was stirred at ambient temperature for 2 hrs. The alkaline mixture was washed with chloroform (400 mL×3) and the aqueous layer was separated and adjusted the pH to 1-2 with 6 N hydrochloric acid. The acidic solution was extracted with chloroform (400 mL×3). The organic layer was separated and dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. Semi-purified (q), purity: 97.7% by HPLC (see Example 25).

(7) Purification of Compound (q) (2)

Semi-purified (q) was dissolved in chloroform (280 mL) and n-hexane (400 mL) was added on the surface of the solution and the resulting mixture was placed at ambient temperature for 15 hrs. The precipitate was removed by filtration and the filtrate was evaporated under reduced pressure to dryness. Compound (q); liver oil, 47.4 g (86% yield), content: 84.5% by HPLC (see Example 25), 99.7% ee by chiral HPLC (see Example 25).

¹H-NMR (400 MHz, CDCl₃) δ: 0.94 (3H, t, J=7.3 Hz, CH₂CH₃ ), 1.75 (2H, q, J=7.3 Hz, CH₂CH₃ ), 3.58 (1H, s, OH), 3.96 (3H, s, OCH₃), 5.16 (1H, d, J=15.6 Hz), 5.47 (1H, d, J=15.6 Hz), 7.59 (1H, s, aromatic-H). [α]_(D) ²⁰=+51.3 (c=0.981, CHCl₃)

(8) Synthesis of Compound (r)

A solution of Compound (q) (42.8 g, 0.10 mol, content: 84.5%), palladium acetate (1.34 g, 5.95 mmol) and potassium carbonate (24.67 g, 0.179 mol) in propanol (490 mL) was degassed by suction and replaced with argon gas and degassed by suction and then charged with carbon monoxide. The mixture was stirred at 60° C. for 4 hrs. After cooling, the insoluble material was removed on a Celite pad and the material on the pad was washed with ethyl acetate (300 mL). The filtrate was washed with 1 N hydrochloric acid (150 mL) and brine (300 mL) and the aqueous layer was separated and extracted with ethyl acetate (300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. The residue was purified through silica gel column chromatography; silica gel (200 g), eluent: chloroform-methanol (99:1). Compound (r): brown oil, 30.3 g (70% yield), content: 73.4% by HPLC (see Example 26).

¹H-NMR (400 MHz, CDCl₃) δ: 0.88 (3H, t, J=7.3 Hz, CH₃), 1.04 (3H, t, J=7.3 Hz, CH₃), 1.82 (4H, m, CH₂×2), 3.69 (1H, s, OH), 4.09 (3H, s, OCH₃), 4.34 (2H, t, J=6.8 Hz, CH₂), 5.31 (1H, d, J=16.3 Hz), 5.61 (1H, d, J=16.3 Hz), 7.94 (1H, s, aromatic-H)

(9) Synthesis of Compound (s)

To a stirred solution of Compound (r) (28.7 g, 68.2 mmol, content: 73.4%) and sodium iodide (27.6 g, 0.18 mol) in absolute acetonitrile (141 mL), chlorotriethylsilane (23.3 mL, 0.18 mmol) was added dropwise at ambient temperature under nitrogen atmosphere. The mixture was stirred at ambient temperature for 3 hrs and then quenched by 1 N hydrochloric acid (8 mL) and 10% sodium sulfite (232 mL). The resulting mixture was stirred at ambient temperature for 30 min. The mixture was extracted with ethylacetate and the organic layer was separated and evaporated under reduced pressure to dryness. Compound (s): 22.3 g (95% yield), content: 85.6% by HPLC (see Example 27).

¹H-NMR (400 MHz, CDCl₃) δ: 1.00 (3H, t, J=7.3 Hz, CH₃), 1.02 (3H, t, J=7.3 Hz, CH₃), 1.83 (4H, m, CH2×2), 3.75 (1H, s, OH), 4.35 (2H, t, J=6.8 Hz, CH₂), 5.21 (1H, d, J=17.1 Hz), 5.61 (1H, d, J=17.1 Hz), 7.28 (1H, s, aromatic-H), 9.59 (1H, brs, OH).

(10) Synthesis of Compound (t)

A solution of Compound (s) (0.50 g, 1.46 mmol, content: 86.6%) and potassium carbonate (0.40 g, 2.92 mmol) in dimethylsulfoxide (7 mL) was stirred at 50° C. under argon atmosphere for 20 min. To the stirred mixture, tert-butyl acrylate (2.1 mL, 14.6 mmol) was added dropwise under argon atmosphere and the stirring was continued for 20 hrs. Water (10 mL) and concentrated hydrochloric acid (1 mL) were added dropwise to the stirred mixture in an ice-cooled bath. The mixture was extracted with toluene-ethyl acetate (4:1, 7 mL×4). The combined extracts were washed with water (5 mL×3), dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure to dryness. Compound (t) 0.55 g (77% yield), content: 75.0% by HPLC (see Example 28).

¹H-NMR (400 MHz, CDCl₃) δ: 0.99 (3H, t, J=7.4 Hz, CH₂CH₃ ), 1.58 (9H, s, t-Bu), 1.83 (2H, m, CH₂ CH₃), 4.68 (2H, s, CH₂), 5.25 (1H, d, J=17.8 Hz), 5.69 (1H, d, J=17.8 Hz), 7.01 (1H, s, aromatic-H).

(11) Synthesis of Compound (h)

To a solution of Compound (t) (1.02 g, 1.84 mmol, content: 66.0%) in toluene (17 mL), trifluoroacetic acid (1.7 mL) was added with stirring under argon atmosphere. The mixture was stirred at 110° C. under argon atmosphere for 100 min. After cooling, the mixture was evaporated under reduced pressure to dryness. The residue was suspended in dichloromethane (50 mL) and the insoluble material was filtered on a Celite pad. The filtrate was washed with water (10 mL) and the organic layer was separated and the aqueous layer was extracted with dichloromethane (20 mL x3). The combined extracts were dried over anhydrous sodium sulfate, filtered and evaporated under reduces pressure to dryness. Compound (h);

(S)-4-ethyl-7,8-dihydro-4-hydroxy-1H-pyrano-[3,4-f]indolidine-3,6,10(4H)-trione; 0.46 g (77% yield), content: 80.7% by HPLC (as follows).

¹H-NMR (400 MHz, CDCl₃) δ: 0.98 (3H, t, J=7.3 Hz, CH₂CH₃ ) 1.81 (2H, m, CH₂ CH₃), 2.97 (2H, t, J=6.3 Hz, CH₂ CH₂), 3.64 (1H, s, OH), 4.34 (2H, m, CH₂CH₂ ), 5.25 (1H, d, J=17.1 Hz), 5.68 (1H, d, J=17.1 Hz), 7.22 (1H, s, aromatic-H).

HPLC Operating Conditions

-   -   Column: Inertsil ODS-2, 0.46 cm ID×25 cm     -   Temperature: constant temperature at around 40° C.     -   Flow rate: 1 mL/min     -   Mobile phase: 10 mM potassium dihydrogenphosphate-methanol (4:1)     -   Detect: 254 nm

EXAMPLE 31 Synthesis of 7-ethyl-10-hydroxycamptothecin (SN-38)

A suspension of Compound (h) (0.50 g, 1.82 mmol, content: 96.6%), obtained as described in Example 30 (11), and Compound (e) (0.36 g, 2.14 mmol) in the presence of p-toluenesulfonic acid monohydrate (10 mg) in acetic acid-toluene (1:1, 10 mL) was stirred at 100° C. under nitrogen gas atmosphere for 18 hrs. The mixture was evaporated under reduced pressure and toluene (10 mL) was added to the residue and then evaporated under reduced pressure to dryness. The residue was suspended in acetone (9 mL) and the suspension was stirred at ambient temperature for 2 hrs. The suspension was filtered by suction and the collected solid was washed with acetone (2 mL×2) and then dried under reduced pressure. SN-38: brown solid, 0.63 g (89% yield), content: 97.7% by HPLC (see Example 9).

¹H-NMR (400 MHz, CDCl₃) δ: 0.98 (3H, t, J=7 Hz, CH₃), 1.38 (3H, t, J=7 Hz, CH₃), 1.90 (2H, q, J=7 Hz, CH₂), 3.08 (2H, q, J=7 Hz, CH₂), 5.17 (2H, s, CH₂O), 5.23 (1H, d, J=16 Hz), 5.54 (1H, d, J=16 Hz), 7.34-7.39 (3H, m), 6.83 (1H, d, J=9 Hz).

EXAMPLE 32 Synthesis of 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (SN-38B-11)

SN-38 (0.91 g, 2.32 mmol), obtained as described in Example 31, was converted into SN-38B-11 (1.22 g, 89% yield, 99.8% ee by the chiral HPLC conditions; see Example 10) by the reported method (S. Sawada, et al., Chem. Pharm. Bull., 1991, 39, 1446).

EXAMPLE 33 Synthesis of 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin hydrochloride trihydrate (CPT-11)

SN-38B-11 (1.00 g, 1.7 mmol), obtained as described in Example 32, was dissolved in 0.1 N hydrochloric acid (20 mL) by heating at around 80° C. Acetonitrile (100 mL) was added to the solution and the mixture was gently stirred at ambient temperature for 15 hrs. The precipitates were filtered by suction and dried under reduced pressure and then humidified. CPT-11: pale yellow crystalline powder, 0.95 mg (89.9% yield).

INDUSTRIAL APPLICABILITY

By use of the synthetic process of the invention highly pure 2′-amino-5′-hydroxypropiophenone and tricyclic ketone can be synthesized in a short time with a high recovery yield, and by use of these as intermediates a total synthesis of CPT analogs can efficiently be carried out. 

1. A process for preparing 2′-amino-5′-hydroxypropiophenone to synthesize camptothecin analogs, wherein from Compound (a):

Compound (b):

is produced by mixing Compound (a), a benzylation reagent and a base, and stirring said mixture in solvent under reflux; and from Compound (b) Compound (c):

is produced by dropping Grignard reagent to Compound (b) under an inert gas atmosphere; and from Compound (c) Compound (d):

is produced by mixing Compound (c) and an oxidizing agent and stirring the mixture; and from Compound (d) Compound (e):

is produced by a catalytic reduction; wherein R is a protective group which can be deprotected by a catalytic reduction.
 2. A process according to claim 1, wherein the protective group which can be deprotected by a catalytic reduction is a benzyl group.
 3. A process according to claim 1, wherein the solvent is dimethylformamide.
 4. A process according to claim 1, wherein the Grignard reagent is vinyl magnesium bromide.
 5. A process according to claim 1, wherein the oxidizing agent is Jones reagent, manganese dioxide, TEMPO-sodium hypochlorite, Dess-Martin Periodinane, PCC, PDC or DMSO/oxalyl chloride/triethylamine (Swern oxidation).
 6. A process for preparing 2′-amino-5′-hydroxypropiophenone to synthesize camptothecin analogs, wherein from Compound (a):

Compound (c″):

is produced by dropping Grignard reagent to Compound (a) under an inert gas atmosphere; and from Compound (c″) Compound (d″):

is produced by mixing Compound c″ and an oxidizing agent and stirring the mixture; and from Compound (d″) Compound (e):

is produced by a catalytic reduction of Compound (d″).
 7. A process according to claim 6, wherein the Grignard reagent is vinyl magnesium bromide.
 8. A process according to claim 7, wherein the oxidizing agent is Jones reagent, manganese dioxide, TEMPO-sodium hypochlorite, Dess-Martin Periodinane, PCC, PDC or DMSO/oxalyl chloride/triethylamine (Swern oxidation).
 9. A process according to claim 1, wherein the benzylation reagent is an aryl halide, phenyldiazomethane or dibenzyl carbonate, and a metal catalyst is used in the catalytic reduction.
 10. A process according to claim 9, wherein the aryl halide is benzyl chloride, benzyl bromide, benzyl iodide, and the metal catalyst is palladium-carbon, palladium hydroxide-carbon or rhodium-alumina.
 11. A process according to claim 10, wherein the oxidizing agent is Jones reagent, manganese dioxide, TEMPO-sodium hypochlorite, Dess-Martin Periodinane, PCC, PDC or DMSO/oxalyl chloride/triethylamine (Swern oxidation).
 12. A process according to claim 6, wherein a metal catalyst is used in the catalytic reduction.
 13. A process according to claim 12, wherein the metal catalyst is palladium-carbon, palladium hydroxide-carbon or rhodium-alumina.
 14. A process according to claim 13, wherein the oxidizing agent is Jones reagent, manganese dioxide, TEMPO-sodium hypochlorite, Dess-Martin Periodinane, PCC, PDC or DMSO/oxalyl chloride/triethylamine (Swern oxidation). 